End users "Feedback" to improve ergonomic design of machinery.

This paper describes the Feedback method designed to collect the contribution of users for the reconstruction and comprehension of the actual work and real activity for the improvement of the technical standards, design, manufacturing and use of machinery. The Feedback method has since now been applied successfully - in collaboration with public authorities, market surveillance bodies, social partners organization and technical institutes - to five different types of machines: woodworking machinery, forklift trucks, angle grinder and combine harvester. After ten years of experimentation in seven European countries Feedback has proved to be trans-nationally comparable and has attracted the interest of as much as 250 expert users - mostly workers, but also employers and technicians - who have shared their knowledge and experience by taking part in almost 30 working groups. The information collected with the Feedback method can be used by: -CEN and ISO standardization committees and working groups to become aware of the problems relating to the real use of specific machines in different work contexts, and thus to be able to draw up new or to revise existing standards accordingly; - Designers and manufacturers to produce better, more comfortable and safer machines and to provide precise instructions for use; - Employers, users and workers for training purposes and for defining appropriate work procedures; - Inspection bodies to enhance their knowledge and improve the efficiency of their interventions and advice.

Geriatric teledermatology: store-and-forward vs. face-to-face examination.

BACKGROUND: Telemedicine could be useful in countries like Italy to meet the needs of elderly patients and in particular in those in precarious general conditions, for whom travelling even short distances can pose considerable practical and economical difficulties. OBJECTIVE: The aim of this study was to determine the efficacy of store-and-forward teledermatology vs face-to-face consultations in elderly patients. METHODS: A total of 130 geriatric patients with skin diseases requiring dermatological examination were enrolled. The patients examined, consisting of 60 men (46.15%) and 70 women (53.85%), were aged between 66 and 97 years (mean age 80.58 years). Three dermatologists of the department, with equal experience took turns in face-to-face examination and teledermatology (store-and-forward). To compare face-to-face dermatological examinations with the asynchronous store-and-forward approach of teledermatology, we considered diagnostic agreement (ICD-9 code), therapeutic agreement and concordance of diagnostic confidence. RESULTS: One hundred and fourteen of 130 patients were diagnosed with the same ICD-9 code, making a total observed agreement of 87.7% with a Cohen's κ estimated of 0.863. Agreement between therapies was 69.6% (Cohen's κ = 0.640). As it concerns diagnostic confidence, dermatologists appeared generally slightly less certain of their diagnosis by telemedicine. CONCLUSIONS: Store-and-forward teledermatology can improve diagnostic and therapeutic care for skin disease in elderly who lack easy and/or direct access to dermatologists.

Acellular pertussis vaccines containing genetically detoxified pertussis toxin induce long-lasting humoral and cellular responses in adults.

New generation pertussis vaccines, containing only purified Bordetella pertussis antigens, have been proven safe, immunogenic and efficacious. They have, however, raised new questions regarding the mechanism of protection from whooping cough and the duration of the immune response following vaccination. In addition to the antibody (Ab) titer, the level of pertussis toxin (PT) neutralizing antibodies may be very important in protection and the role of cell-ediated immunity needs to be defined. We have previously reported the safety and immunogenicity results of two phase I trials in adult volunteers with two acellular pertussis vaccines containing genetically detoxified PT alone or in combination with filamentous hemagglutinin (FHA) and 69K protein. In this work, we present the results of a long term follow-up study of the immune response in the same vaccinees. We evaluated the Ab response, the PT neutralizing titer and the peripheral blood T cell response up to 4 years following vaccination. Our results show that in adults the level of antibodies to PT, FHA and 69K and the PT neutralizing titers slightly decline between 2.5 and 12 months after the last vaccine dose, but they remain high in the following 2-4 years, showing levels 10-100 times higher than pre-vaccination values. The T cell responses were more heterogeneous among vaccinees but they did not show any significant decline throughout the period monitored.

Immunogenicity of an acellular pertussis vaccine composed of genetically inactivated pertussis toxin combined with filamentous hemagglutinin and pertactin in infants and children.

We studied the immunogenicity of an acellular pertussis vaccine composed of genetically detoxified pertussis toxin (PT-9K/129G), filamentous haemagglutinin, and a 69-kilodalton protein, pertactin, in 30 children aged 12 to 24 months and in 80 infants aged 2 to 4 months. A significant increase of the neutralizing titer and of the titers against pertussis toxin, filamentous hemagglutinin, and pertactin, as determined by enzyme-linked immunosorbent assay, was achieved after three doses of vaccine in all the children; a significant increase of these antibody titers was obtained in 100%, 96.1%, 93.5%, and 98.7% of the infants, respectively.

Cellular pertussis vaccine containing a Bordetella pertussis strain that produces a nontoxic pertussis toxin molecule.

Bordetella pertussis 165-9K/129G, which produces a nontoxic form of pertussis toxin (PT), was used to prepare a whole-cell diphtheria-tetanus-pertussis (DTP) vaccine. The in vivo potency and the serological response induced by this vaccine were comparable to those of the conventional DTP vaccine which contains active PT. The toxic activities induced by PT such as leukocytosis, histamine sensitivity, and potentiation of anaphylactic reactions, which are present in the conventional DTP vaccine, were absent in the new vaccine. These results suggest that the introduction of a whole-cell vaccine containing B. pertussis 165-9K/129G would induce the same immunity as the conventional vaccine and would avoid the administration of a harmful toxin to children.

Differential activity of interleukin 1 alpha and interleukin 1 beta in the stimulation of the immune response in vivo.

The biological activities of human recombinant interleukin (IL) 1 alpha and IL 1 beta were compared in different biological systems. The two IL 1 forms were equally active in vitro in inducing proliferation of murine thymocytes and of the murine T helper clone D10.G4.1, and in triggering release of prostaglandin E2 from human skin fibroblasts. In vivo, IL 1 alpha and IL 1 beta were similarly pyrogenic both in rabbits and mice, and could equally increase the circulating levels of the acute phase protein serum amyloid A in mice. However, only IL 1 beta showed immunostimulatory activity in vivo, as it could enhance the number of specific antibody-producing cells in the spleen of mice immunized with either a T-dependent or a T-independent antigen. Although devoid of immunostimulatory activity, IL 1 alpha could efficiently compete immunostimulation induced by IL 1 beta, suggesting an effective interaction with the IL 1 receptor. Thus, IL 1 beta appears to have an important role in the positive regulation of immune responses, while IL 1 alpha may act as down-regulator of the IL 1 beta effect.

Radiolabeling of the biologically active peptide 163-171 of human interleukin-1 beta.

N-succinimidyl[2,3-3H]propionate was used for the radiolabeling of the biologically active peptide fragment 163-171 of human interleukin-1 beta (VQGEESNDK). Suitable reaction conditions were studied to obtain useful labeling of the molecule. A mixture of mono- (70%) and bi- (30%) propionyl derivatives was obtained with a total 3H specific activity of 87 Ci/mmol of peptide. The conditions for an efficient chromatographic separation of labeled peptide from unreacted reagents and by-products were established. The labeled peptide maintained the same biological activity as that of the corresponding unlabeled molecule, indicating that the labeling procedure did not alter the biological characteristics of the peptide. This thus allows the use of the radiolabeled peptide for receptor binding studies.

Differential binding of IL-1 alpha and IL-1 beta to receptors on B and T cells.

The interleukin 1 receptors (IL-1R) on the human B lymphoma RAJI and on the murine thymoma EL4-6.1 have been characterized. Equilibrium binding analysis using both 125I-labeled IL-1 alpha and IL-1 beta showed that RAJI cells have a higher number of binding sites/cell for IL-1 beta (2400, Kd 2.2 nM) than for IL-1 alpha (316, Kd 0.13 nM). On the other hand, EL4-6.1 cells have more receptors/cell for IL-1 alpha (22 656, Kd 1 nM) than for IL-1 beta (2988, Kd 0.36 nM). Dexamethasone (DXM) induced on RAJI cells a time-dependent increase in binding sites for both IL-1 beta and IL-1 alpha without affecting their binding affinities. However, while receptor-bound 125I-IL-1 alpha was displaced with equal efficiency by both IL-1 forms, only unlabeled IL-1 beta could effectively displace 125I-IL-1 beta. Cross-linking experiments indicated that RAJI cells have a predominant IL-1R of about 68 kDa, while EL4-6.1 cells have an IL-1-binding polypeptide of 80 kDa. These results suggest that B and T cells possess structurally different IL-1R with distinct binding properties for IL-1 alpha and IL-1 beta.

Interleukin 1 stimulates production of LTC4 and other eicosanoids by macrophages.

Human recombinant IL-1 alpha stimulates in a dose-dependent fashion the production of 5-lipoxygenase-derived LTC4 and, to a lesser extent, of cyclooxygenase-derived eicosanoids (PGE2, 6-keto PGF1 alpha, TXB2) by murine resident peritoneal macrophages. Enhancement of eicosanoid production was evident on unstimulated macrophages, but only marginal on macrophages phagocytosing zymosan. The effect of IL-1 was similarly achieved at physiological (37 degrees C) as well as at higher (39 degrees C) temperatures. Human recombinant IL-1 beta, human natural IL-1, and partially purified murine IL-1-rich supernatant also stimulated eicosanoid production by macrophages, although IL-1 alpha appeared to be the most effective.

Mapping of biologically relevant sites on human IL-1 beta using monoclonal antibodies.

mAb have been raised that recognize human IL-1 beta. Using overlapping peptide fragments expressed in yeast and bacteria, we have mapped the regions of the protein to which these antibodies bind. To assess the relevance of the different regions of IL-1 beta for the expression of its biologic activity, the ability of the antibodies to block IL-1 activity was assayed. Antibodies recognizing the regions 133-148 and 251-269 of human IL-1 beta could inhibit the activity of IL-1 beta, but not of IL-1 alpha, in two different biologic assays, the murine thymocyte proliferation and PGE2 release from human fibroblasts. Conversely, antibodies that recognize the region 218-243 have only a moderate inhibitory effect on the IL-1 beta biologic activity in both assays. Finally, an antibody mapping to the region 148-192 did not inhibit IL-1 beta activity either on thymocytes or on fibroblasts. It is suggested that IL-1 beta-induced cell activation involves different regions of the protein and that both N-terminal and C-terminal fragments are involved in the correct functioning of the IL-1 beta molecule.

Interferons inhibit LTC4 production in murine macrophages.

Mouse resident peritoneal macrophages (M phi) produce the highly bioactive eicosanoid LTC4 when stimulated in vitro with zymosan or with the calcium ionophore A23187. This production was dramatically inhibited in M phi pre-exposed to IFN-alpha, IFN-beta, or IFN-gamma. Although all IFN were able to decrease the availability in M phi of the LTC4 precursor AA, this decrease was not the only cause of the IFN-induced inhibition of LTC4. In fact, further analysis of the different steps of the LTC4 biosynthetic pathway revealed that IFN-gamma could inhibit the formation of LTA4, thus of its derivatives LTC4 and LTB4, possibly acting at the level of the enzyme LTA4-synthetase. In contrast, IFN-alpha and IFN-beta only depressed the ability of M phi to metabolize AA into LTC4, leaving unaltered the synthesis of LTB4. However, IFN-alpha and IFN-beta did not influence directly the activity of any of the enzymes involved in LTC4 biosynthesis, indicating that they may act through some indirect, as yet unidentified regulatory mechanism. These data suggest that IFN-alpha and IFN-beta and, in different situations, IFN-gamma can be potentially useful in vivo in antagonizing localized anaphylactic or inflammatory reactions.

Anti-inflammatory activity of IFN-beta in carrageenan-induced pleurisy in the mouse.

The effect of IFN-beta on the development of the inflammatory reaction was studied in an experimental animal model, carrageenan-induced pleurisy in the mouse. Intrapleural inoculation of IFN-beta at the same time as carrageenan administration inhibited both migration of inflammatory cells and exudate formation in the pleural cavity in a dose-dependent fashion. Similarly, IFN-beta decreased the presence of the arachidonate metabolites PGI2, TXA2 and PGE2 (highly active molecules involved in the regulation of the inflammatory reaction) in inflammatory exudates. A marked inhibition of the inflammatory response to carrageenan was also evident when IFN-beta was administered several hours after the inflammatory challenge. In contrast, administration of IFN-gamma did not modify significantly any of the inflammatory parameters considered.

Regulation of arachidonic acid metabolism in macrophages by immune and nonimmune interferons.

Mouse resident peritoneal M phi release AAA and metabolize it into cyclooxygenase- and lipoxygenase-derived eicosanoids, when triggered in vitro with different stimuli. Pretreatment of M phi with nonimmune IFN-alpha and IFN-beta dramatically decreased AA liberation from M phi phospholipids and eicosanoid formation after stimulation of M phi with Zy, A23187, or PMA. M phi exposed to immune IFN-gamma also showed a substantial impairment of both AA liberation and eicosanoid production upon exposure to Zy. However, AA and eicosanoid release was increased by IFN-gamma, rather than depressed, in PMA-triggered M phi. In addition, IFN-gamma showed differential effects on M phi stimulated with A23187. In fact, it inhibited AA release as well as formation of lipoxygenase-derived LTC4, but it highly increased the release of the cyclooxygenase products PGE2 and 6-keto PGF1 alpha. The ability of IFN-gamma to differentially modulate AA metabolism of M phi, depending on the nature of the triggering agent, sets forth the high specificity of the regulatory capacity of this molecule. This is at variance with the down regulation of AA metabolism that is generally observed with nonimmune IFN.

Interferon inhibits prostaglandin biosynthesis in macrophages: effects on arachidonic acid metabolism.

Mouse resident peritoneal M phi produced considerable amounts of PGE when triggered in vitro with soluble or particulate stimuli. Preexposure of M phi to IFN-beta dramatically decreased PGE production. This effect depended on the dose of IFN-beta used and was abolished by anti-IFN-beta globulin. In addition to PGE, other AA metabolites of the cyclooxygenase pathway, namely TXB2 and 6-keto PGF1 alpha, were decreased in IFN-beta-treated M phi. However, IFN-beta did not have any effect on cyclooxygenase activity of M phi, indicating that an earlier step of PG biosynthesis was likely to be the target of its inhibitory action. Indeed, the release of radioactive compounds from M phi prelabeled with [14C]AA was strongly impaired by IFN-beta, suggesting that IFN-beta would decrease PG production in M phi by blocking phospholipase activation. The possibility that the IFN-beta effect on phospholipase could be mediated through the increase of the intracellular levels of cAMP is suggested.

Dissociation between macrophage tumoricidal capacity and suppressive activity: analysis with macrophage-defective mouse strains.

Macrophages (M phi diameter) from three mouse strains with genetically distinct M phi diameter deficits (C3H/HeJ, A/J, and P/J) were unable to develop high cytolytic and cytotoxic activity against tumor cells in vitro when exposed to agents (MAF and IFN-beta) that strongly increased the tumoricidal capacity of M phi diameter from nondefective C3H/HeN mice. Nevertheless, the tumoricidal deficits of M phi diameter from the defective strains did not affect their suppressive capacity on Con A-induced lymphoproliferation, nor their ability to react to IFN-beta by decreasing suppressive activity. In fact, natural suppressive activity and IFN-beta-induced changes in the suppression of M phi diameter from C3H/HeJ, A/J, and P/J mice were highly comparable to those of C3H/HeN M phi diameter, thus stressing the dissociation between the mechanisms governing M phi diameter suppression and M phi diameter tumoricidal activity. Analysis of the modulation by MAF and IFN-beta of M phi diameter ability to release the oxygen metabolites O2- and H2O2, molecules possibly involved in the effector mechanism of both M phi diameter cytotoxicity and suppression, revealed a close correlation with the patterns of suppressive activity in both nondefective and defective strains. In contrast, no correlation between the production of oxygen-reactive species and M phi diameter tumoricidal activity was observed. The ability of MAF- and IFN-beta-treated M phi diameter to produce PGE, a molecule of major importance in M phi diameter-mediated suppression and possibly involved also in the regulation of M phi diameter tumoricidal activity, again paralleled M phi diameter suppressive capacity. Thus, the mechanisms controlling M phi diameter antitumor activity appeared to be clearly distinct from those involved in M phi diameter suppression.