[A new method of functional rehabilitation of patients with occupational disability caused by lumbar spine pathology]. / Un nuovo metodo di riabilitazione funzionale (DBC) in pazienti con disabilità lavorativa da patologia del rachide lombare.

Rehabilitation is reducing disability through an improvement of function, i.e. rehabilitation need always to be "functional". Nevertheless, like it happened in other fields where rehabilitation has not yet reached its maturity, in spinal pathologies it has usually been considered equal to conservative treatment, i.e. not surgical relief of pain. But relieving pain does not always mean to recover function: in fact, it has been proven that there are no long term good results treating symptomatically chronic low back pain (CLBP). In the Eighties in USA a new kind of treatment has been introduced for inpatient rehabilitation of CLBP, based on a full-time four-weeks training of physical abilities, together with a cognitive and educational approach to pain. This treatment, named "functional rehabilitation", proved its efficacy also in randomised controlled trials (RCT). In the Nineties in Finland the outpatient version of this treatment for CLBP has been developed by "Documentation Based Care" (DBC), whose efficacy have also been documented in RCT: the efficiency of this last kind of treatment should be higher. DBC treatment is spreading around the world, and it recently reached Italy in Don Gnocchi.

Use of an enzyme-linked immunosorbent assay for screening hybridoma antibodies against hepatitis B surface antigen.

An enzyme-linked immunosorbent assay (ELISA) was developed for screening production of monoclonal antibodies with specificity for HBsAg. Mouse hybridoma IgG were firstly extracted from assay medium with goat anti-mouse IgG adsorbed on polystyrene beads. The specific antibody was revealed by saturation with HBs antigen from human positive sera followed by reaction with specific sheep anti-HBs antibody conjugated with peroxidase. The sensitivity was of the order of 0.5-1 ng/ml specific antibody and specificity was satisfactory. The assay permits easy identification of determinant specificity on screening.

ELISA for specific anti-toxoplasma IgM antibodies: aspects related to serum interference.

Two different methods were used to prepare solid-phase antigen (Ag) from soluble extracts of tachyzoites of Toxoplasma gondii: (A) physical adsorption on polystyrene beads; and (B) formaldehyde fixation of Ag previously dried in microtitration wells. In both cases a horseradish peroxidase conjugate with anti-IgM IgG was used as tracer. The assay scheme consisted of sequential incubations of diluted serum samples and tracer solution (1 or 2 h, 37 degrees C), colour development in the presence of substrate (10 min at room temperature), addition of H2SO4, and absorbance reading at 492 nm. In procedure A no cut-off value for positives could be determined owing to a large overlap between positive and negative sera. The extent of overlap directly correlated with the total IgM content of samples. With negative sera similar values were obtained with sensitized and untreated beads: thus a correction could be made by directly subtracting absorbance values determined in parallel runs with uncoated beads. Results with negative sera correlated with total IgM concentration in procedure B also, but much less variability of blank values allowed negative and positive sera to be effectively discriminated. A series of reference positive and negative sera was correctly classified by both procedures A and B. However, the latter appeared preferable, as not requiring blank correction.

ELISA for antibody measurement: aspects related to data expression.

In an ELISA for antitoxoplasma IgG (antigen-coated polystyrene beads, horseradish peroxidase-coupled IgG or staphylococcal protein A), 3 modes of expressing the analytical results were considered, i.e. end-point antibody titre, untransformed absorbance reading at a single sample dilution, and antibody-activity unit from a calibration response curve (reference sera as calibrators). Criteria of merit for evaluation were defined as (a) stability of data under various conditions relating to both changes in assay design and minor variability of experimental conditions, and (b) linearity of response with dilutions of positive sera in negative serum, i.e., with positive sera sequentially defined as to antibody activity. Conclusions emerging were: single absorbance readings have some validity as indicators of trends but are very prone to systematic and random variability and inconsistent in response-antibody activity parallelism; parallelism of response proved to be an advantage of titration, but disadvantages are lack of practicability (manipulation and reagent costs involved) and lack of reliability (high levels of systematic and statistical error) The introduction of a reference scale allowing data to be expressed in activity units eliminates systematic components of error and gives analytical consistency. Use of the latter appears mandatory for between-run, between-laboratory and between-method normalization.

ELISA for toxoplasma antibody detection: a comparison with other serodiagnostic tests.

An ELISA method was developed for the measurement of toxoplasma IgG antibodies in human serum using antigen-coated polystyrene beads as a solid phase and anti human IgG-horse radish peroxidase conjugate as an enzymatic tracer. In order to assess ELISA sensitivity and specificity, a between methods comparison was made using 'conventional' serological tests as reference (dye-test, crossover-linked immunoassay, passive haemagglutination, indirect immunofluorescence). From an analysis of the group classifications obtained some considerations emerged: the ELISA specificity looks comparable with that of the 'reference' tests, as no sample classified as negative by all these tests was ELISA-positive, and vice versa; ELISA appears to correlate better with haemagglutination and immunofluorescence, on the basis of the respective class frequencies; in particular, the number of positives, which is much lower for the dye-test and crossover-linked immunoassay, suggests that a higher sensitivity is reached in the former cases.

The "soluble sandwich" approach for immunoassays: methodological and instrumental implications.

The simultaneous presence of homogeneous and heterogeneous reactions at different binding sites of a multiepitope antigen makes the description of the kinetic parameters of the so called "one step" solid phase immunometric assays complex. The authors extended the "one step" approach to the concept of the "soluble sandwich" methodology which differs from the former by the delayed solid phase capture of the biotinylated immunocomplex to a streptavidin coated solid support. Using prolactin monoclonal IEMAs as a model, the equilibria involved in the reactions have been studied on a thermodynamic basis through a description of the kinetics of the interactions between biotinylated Mabs and solid phase streptavidin both in presence and/or in absence of the antigen and HRP-conjugated antibody. A comparative evaluation of models in which the biotinylated antibody was previously insolubilized on the streptavidin solid phase has been performed as well. The experimental work was carried out by using 125I labelled McBiot and Prolactin to trace individual interactions and peroxidase/H2O2/TMB systems to develop the enzymatic analytical signals. A new instrument/data reducing system was also optimized to expand the OD reading range provided by conventional, single wavelength colorimeters. The greater flexibility theoretically expected for the "soluble sandwich" approach and the possibility to extend the analyte working range without detrimental effects on the readability of low doses responses have been experimentally confirmed.

Methodological approach and diagnostic usefulness of a new assay for anti-thyroid peroxidase autoantibodies.

Although thyroid microsomal antibodies (anti-MAb) have been recently proven to be directly to thyroid peroxidase (TPO), current methods for MAb detection still employ unpurified microsomal fractions. The authors have therefore developed and evaluated a specific radioimmunoassay (RIA) for autoantibodies to TPO (anti-TPO Ab) based on competitive inhibition of 125I-TPO to an anti-TPO monoclonal antibody coated on plastic microtiter wells (RIA-1) or tubes (RIA-2). Preliminary experiments showed that both assays were able to specifically detect anti-TPO Ab, while negative results were obtained with normal sera and with sera containing other organ- and non-organ-specific autoantibodies including anti-thyroglobulin antibodies (anti-TgAb). No significant difference in sensitivity, specificity and reproducibility was found between RIA-1 and RIA-2, and the results obtained with the two techniques were therefore pooled together. Anti-TPO Ab were then assayed in 110 normal controls and in 441 patients with different autoimmune (AITD) or non-autoimmune (N-AITD) thyroid diseases and compared to anti-M Ab as assessed by passive hemagglutination (PH). Positive anti-TPO Ab were observed in 4/110 (3.6 p. cent) normal controls, 88/117 (80 p. cent) patients with Graves' disease, 122/123 with Hashimoto's thyroiditis or idiopathic myxedema and 21/201 (10.4 p. cent) with miscellaneous N-AITD. A highly significant positive correlation (r = 0.91, p less than 0.0001) was found between anti-MAb by PH and anti-TPO Ab by RIA ;discrepant results were limited to sera with negative or low (1/100-1/400) anti-M Ab titers.(ABSTRACT TRUNCATED AT 250 WORDS)

The validity of clinical examination in adolescent spinal deformities.

STUDY DESIGN: retrospective study on the accuracy and reliability of two clinical tests for scoliosis in young patients. AIM: to evaluate the inter-observer reliability of three non-invasive clinical measurements: hump height (HH), axial trunk rotation (ATR), and distance of the spinous process from the plumb line (DP) in standing; to compare these results with the corresponding radiographic measurements, the Cobb angles (CA). POPULATION: 116 patients, 78 females and 38 males; 410 examinations have been performed (144 patients with brace and 266 without). METHODS: a database was created using the measurements of different clinical parameters obtained from two examiners that measured them independently and in the same conditions. The Cobb method has been used as a gold standard. RESULTS: our results show a very high inter-rater reliability for HH and ATR measurements. The DP shows a different inter-rater reliability for the thoracic (C7) and lumbar (L3) spine, in both cases lower than that in the frontal plane; the ICC of the thoracolumbar DP (D12) was very low. The correlation with the radiographic value was weak.

8-[C]Benzylaminopurine Translocation in Phaseolus vulgaris.

[8-(14)C]Benzylaminopurine (BA) translocation was studied in whole plants of Phaseolus vulgaris L. under three different light regimes (continuous light, 8-hour light + 16-hour dark, dark). Applications were made to the apex, to a cotyledonary leaf, or to the root system. Results showed that no BA basipetal translocation occurred, however BA is easily absorbed by the root system and is translocated acropetally.The amount of BA absorbed and its acropetal translocation rate depend on the light regime. The hypothesis of a passive cytokinin transport through the xylem, regulated by the transpiration stream, is discussed.

[Immobilization of antibodies against Australia antigen on nylon spheres: use in immunoenzymatic determination of HBsAg]. / Immobilizazione di anticorpi anti-antigene Australia su sfere di nylon: utilizzazione in un dosaggio immunoenzimatico di 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.

Rapid detection of antibodies to infectious bovine rhinotracheitis by 'macro' and 'micro' ELISA.

Two kinds of enzyme-linked immunosorbent assay were evaluated in their ability to detect specific antibodies against Bovine Rhinotracheitis Virus (IBR-IPV). The tests were called MACROELISA and MICROELISA, according to the kind of the solid support used for antigen insolubilization, polystyrene beads and microtitration plates respectively. Partially purified virus was used to coat both beads and plates; a single dilution of examples was tested and protein A linked to peroxidase was employed as enzyme tracer. Quantitative instrumental results from MACROELISA and qualitative visual results from MICROELISA were compared with serum neutralization titers. The results clearly show that ELISA tests are suitable for IBR serologic detection, being sensitive, specific and accurate over serum neutralization method.

HBeAg/anti-HBe determination with a new monoclonal immunoradiometric assay.

The performance of two assays for the HBeAg/anti-HBe system associated with hepatitis B virus infection, using monoclonal (EBK-Sorin) and polyclonal (HBe-Abbott) antibodies, has been compared. The results on a random population (1,000 samples) demonstrated for the monoclonal reagent a higher sensitivity, without any loss in specificity, and with the further advantage of the use of lower radioactivity levels. The proportion of positives and negatives obtained with the two kits was found to remain unchanged in the case of HBeAg, while a markedly larger percentage of positives (7% higher) was detected for anti-HBe using the monoclonal antibodies.