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Biomedical subjects

L Callegaro

Publications and source records attributed to L Callegaro.

49 records · Page 3Linked to original sources
PubMed1983

Hollow fiber immobilized L-asparaginase: in vivo and in vitro immunological studies.

The enzyme L-asparaginase was covalently immobilized on the inner surface of the hollow fibers utilized in a commercially available dialyzer by the periodate method. After sterilization with gamma radiation the bioreactor was able to metabolize in vivo, 90 per cent of circulating asparagine in two hours. The absence in blood of asparaginase-related protein fragments, released from the hollow fiber immobilized enzyme, was monitored using a specific enzyme-linked immunosorbent assay (ELISA).

Antibodies↗
PubMed1981

[Immobilization of antibodies against Australia antigen on nylon spheres: use in immunoenzymatic determination of HBsAg].

Antibodies against HBsAg were immobilized by covalent linkage on nylon beads to be used in solid phase enzyme-linked immunoassays. The covalent bond between the protein and the partially hydrolyzed support was obtained via glutaraldehyde or succinylester methods. These particular solid phases were compared, in a conventional enzyme-linked immunosorbent assay for hepatitis B antigen, with polystyrene beads coated by simple adsorption of IgG on plastic surface. The analytical response was more satisfactory when the polystyrene beads were used as immunoextractive support even if more IgG molecules could be immobilized on nylon beads.

Antibodies, Viral↗
PubMed1981

Immobilization of arginase on hollow fiber hemodialyzer.

The enzyme arginase, purified from bovine liver, was covalently immobilized by the glutaraldehyde method to the inner surface of Cuprophan hollow fibers of a conventional hemodialyzer with a surface 1.3 m3. The yield of the process was 0.3 microgram/cm2 of active enzyme at physiological pH. The immobilization method did not adversely affect the physical and mechanical properties of hollow fibers neither their hemocompatibility. After sterilization with ethylene oxide, the bioreactor was able to metabolize five liters of 50 microM arginine solution at pH 7.4, in six hours.

Arginase↗
PubMedDate not supplied

Purification and characterization of Fab fragments from anti-mouse NGF polyclonal antibodies.

A functional role for Nerve Growth Factor (NGF) in the peripheral nervous system is well-documented, but a similar case for NGF in the central nervous system remains to be established. One approach to answering this question would be the availability of high-affinity monospecific Fab fragments obtained against NGF. In the present studies we describe the preparation and characterization of such Fab fragments from anti-mouse NGF polyclonal antibodies. Following their purification by the use of a NGF Sepharose-coupled affinity column, the Fab fragments were examined for biological competence in several ways. In vitro, the anti-Fab fragments blocked the neuronotrophic activity of NGF, as measured by the survival of chicken embryonic day 8 dorsal root ganglion neurons. In vivo, these Fab fragments, when administered systemically to neonatal rats, produced a decrease of noradrenaline levels in two sympathetically innervated organs, the heart and the spleen. These findings suggest that affinity purified Fab fragments of anti-NGF antibodies can be a useful tool for studying the physiological function of NGF in the nervous system.

Animals↗
PubMedDate not supplied

Production and characterization of antibodies to mouse scrapie-amyloid protein elicited by non-carrier linked synthetic peptide immunogens.

Two polyclonal antibodies were raised by immunizing rabbits with two non carrier-linked synthetic peptides whose amino acid sequences corresponded to codons 89-107 (peptide P1) and 219-233 (peptide P2) of the translated cDNA sequence of murine PrP protein. These free peptides, whose structural characteristics in solution were studied by circular dichroism, elicited a reasonable immunologic response in animals. Both antibodies still recognized the corresponding immunogens after affinity chromatography purification. However, only antibodies raised to the former sequence reacted by immunoblot with a purified preparation of murine scrapie amyloid protein. These findings are discussed together with their correlation to peptide structure and the effectiveness of this simplified immunization procedure.

Amino Acid Sequence↗
PubMedDate not supplied

Competitive binding assay for quantitative determination of GM1 ganglioside in plasma and cerebrospinal fluid.

A competitive binding assay for the quantitative determination of GM1 ganglioside is described. After extraction from biological fluids, GM1 was incubated with a known amount of cholera toxin B-subunit conjugated with horseradish peroxidase, and exposed to GM1 adsorbed onto polystyrene microwells. Since GM1 in solution blocks the binding of toxin B-subunit to GM1 adsorbed onto the solid phase, enzyme activity serves as a reciprocal measure of GM1 concentration in the sample. The assay was used to determine the basal level of GM1 in plasma and cerebrospinal fluid in different populations.

Adult↗
PubMedDate not supplied

Radioimmunodetection of melanoma: preliminary results of a prospective study.

A prospective study to evaluate the clinical usefulness of radioimmunodetection of melanoma in clinical practice is ongoing at the National Cancer Institute of Milan, Italy. Technical conditions for the application of the method were previously reported. In this trial, 99mTc-labelled F(ab')2 fragments of the 225.28S monoclonal antibody were used against a high molecular weight melanoma associated antigen (HMW-MAA). Retrospective studies on radioimmunodetection of melanoma have already been made by our group and by other Centers in about 300 patients. This study concerns the evaluation of the regional extension of primary melanoma. 23 patients with 32 suspected lymphatic involvements of melanoma on the trunk and arms underwent immunoscintigraphy. No false positive results were observed; 3 false negatives, one corresponding to a micrometastasis, were noticed. Specificity corresponds to 100% and sensitivity to 78.6%.

Adolescent↗
PubMedDate not supplied

Alpha-fetoprotein monoclonal assay: preliminary clinical findings in a high risk population.

A two-site solid phase immunoradiometric assay was developed for measurement of human alpha-fetoprotein, utilizing two high-affinity monoclonal antibodies directed against distinct and separate epitopes on the proteic structure. The analytical sensitivity of the assay is 0.5 ng/ml. The clinical sensitivity was evaluated by comparison of patients with cirrhosis and patients with hepatocellular carcinoma with cirrhosis. This assay gave good diagnostic discrimination. In a preliminary clinical trial, the specificity of the assay was 92.3%, the clinical sensitivity 88.2%, and predictive values were 78.9% in the clinically positive stage and 96.0% in the negative stage. The diagnostic efficacy of the assay was 91.3%.

Animals↗
PubMedDate not supplied

[Prevalence of Chlamydia pneumoniae in respiratory infections in children: an ambulatory diagnostic problem].

It has been recently suggested that Chlamydia Pneumoniae infection is a common finding among children with acute respiratory diseases. Chlamydia cell culture is difficult and time-consuming to perform. Polymerase chain reaction (PCR) is a more rapid but also more expensive technique used to identify Chlamydia in pharyngeal swab, but it can be performed only in few specialized laboratories. We tested a rapid enzyme immuno-assay to detect Chlamydia in 20 children with respiratory infections (mean age 3.29 years; male:female ratio = 12:8) and in 21 healthy children (mean age 4.70 years male:female ratio = 15:6). Prevalence of Chlamydia isolation from pharyngeal swab was very high in both patients and healthy children without a significative difference in the two considered groups (45% vs 42%, p = 0.8). Specific Chlamydia IgG antibodies were undetectable in all patients and healthy children. Nine out of 20 patients affected by acute respiratory disease were Chlamydia-positive and 11 out 20 were Chlamydia-negative: these two groups didn't differ in regard to clinical and laboratory features, whereas duration of symptoms was significantly longer in Chlamydia-positive patients (9.3 vs 5.5 days, p = 0.014). Our study suggests a high prevalence of Chlamydia pharyngeal swab positivity in both healthy and sick children. Diagnosis of Chlamydia infection was not feasible on the basis of the considered clinical and laboratory findings.

Acute Disease↗
PubMedDate not supplied

Peripheral nerve regeneration through a novel bioresorbable nerve guide.

A nerve guide made of a benzyl ester of hyaluronic acid (HYAFF11p75) was used to bridge 8 mm gaps in rat tibial nerves. Histologic observations indicated that this biomaterial provoked only a transient, modest inflammatory response, and the resorption rate was compatible with the nerve regeneration processes. Phagocytosis of the biomaterial began after neoangiogenesis and cell migration had taken place from both stumps into the nerve guide material. For comparison, the regeneration achieved was evaluated in nerve guides made of either HYAFF11p75 or Silastic, and in nerves repaired with the autograft technique. Recovery was assessed in vivo 90 days after implantation by measuring the nerve compound action potential (CAP) and conduction velocity (NCV) of the regenerated tibial nerve. The results demonstrate that the nerve guide tubes made of HYAFF11p75 were able to support and direct axonal growth, thereby suggesting a possible use for such biomaterial in the management of short nerve gaps in human pathology.

Animals↗