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L'Italia, insieme al belgio, è uno dei pochi Paesi europei che, dopo il recepimento della Direttiva Europea 2001/20/EC, ha elaborato una normativa ad hoc sugli studi no-profit. Dopo sei anni dall'entrata in vigore del Decreto Ministeriale del Dicembre 2004, sulla base del loro andamento, dell'esperienza diretta del Comitato Etico dell' AUSL di Bologna, dei dati della Regione Emilia-Romagna e dell'Osservatorio nazionale sulla Sperimentazione Clinica dei medici dell'AIFA (OsSC), viene valutato l'impatto della normativa sull'attività dei Comitati Etici e sulla Ricerca in Italia. Si propongono inoltre alcune riflessioni come spunto per la discussione, in previsione di un aggiornamento della normativa di riferimento dei Comitati Etici. http://www.dialogosuifarmaci.it/

Human neutrophils were isolated both from peripheral blood (PB) and from aseptic inflammatory exudates obtained by the Senn's skin window technique (SW). The respiratory burst (O2– production) induced by in response to n-formyl-methionyl-leucyl-phenylalanine (fMLP) and by serum-treated zymosan (STZ) was investigated using a microplate assay. SW neutrophils were primed to enhanced fMLP-dependent O2– production in response to fMLP but not to STZ. Pentoxifylline, a cAMP-elevating drug, dose-dependently inhibited the respiratory burst in any experimental condition, but the dose-effect curves were markedly different according the stimulant used and the source of the cells. With fMLP as stimulant, a significant inhibition of the O2– production by PB neutrophils was obtained using doses of 10 μg/ml, while SW neutrophils were inhibited only by doses equal or higher than 100 μg/ml. With STZ as stimulant, the inhibition of the respiratory burst of PB neutrophils and of SW neutrophils was obtained only with doses higher than 400 μg/ml and 1 mg/ml respectively. Pentoxifylline dose-dependently (10 μg/ml to 1 mg/ml) increased the intracellular adenosine 3′–5′-cyclic monophosphate (cAMP) to the same extent in SW and in PB neutrophils. These data indicate that the priming of neutrophil oxidative metabolism by in vivo inflammation is associated with an increase in the resistance to the regulating effect of cAMP on the fMLP-dependent activation pathway of NADPH oxidase. The fact that therapeutic doses of pentoxifylline do not inhibit the respiratory burst of primed neutrophils may have relevance in the interpretation of the clinical effects of this drug.

Neutrophils, treated with sequential additions of bacterial products such as endotoxin (E. Coli lipopolysaccharide, LPS) and the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP), undergo to metabolic activation and express membrane-anchoring proteins that promote adhesion to serum-coated culture wells. By investigating the dose–response relationships of these phenomena, we have found that: (a) resting neutrophils do not produce a significant amount of superoxide (O) and show only minimal adhesion to serum-coated plastic surfaces; (b) fully activatory doeses (> 5 × 10−8M) of fMLP induce the release of O and a significant increase of the cell adhesion; (c) pretreatment of the cells for 1 h with LPS augments cell adhesion to serum-coated culture wells in the absence of further stimulation and primes the neutrophils to enhanced fMLP-dependent O release; (d) addition of low, substimulatory doses of fMLP (from 10−10M to 5 × 10−9M) inhibits and reverses the adhesion of LPS-treated cells, (e) high fMLP doses (> 10−7M) are additive to LPS in promoting adhesion. Phorbol-myristate acetate (> 10−9M) increased adhesion in both normal and LPS-treated neutrophils, but low doses of this stimulant did not inhibit adhesion. Low doses (10−9M) of fMLP increased intracellular cyclic AMP in both normal and LPS-treated neutrophils, suggesting that stimulus-induced rises in cAMP may be the negative signal responsible for down-modulation of adhesion. Low (5 × 10−9M) and high (5 × 10−7M) fMLP doses induced the same increase of expression of CD11/CD18 integrins, indicating that the inhibition of adhesion caused by low doses is not due to quantitative down-regulation of integrins. These findings may provide an in vitro model of the complex biological events involved in the regulation of neutrophil adhesion.

An objective approach for monitoring the treatment of acute pulmonary exacerbation in cystic fibrosis was evaluated. Eleven biochemical markers of inflammation (erythrocyte sedimentation rate, neutrophil count, C-reactive protein, alpha-1 antitrypsin, haptoglobin, ceruloplasmin, fibronectin, alpha-1 glycoprotein, alpha-2 macroglobulin, C3, granulocyte elastase and anti-Pseudomonas IgG) were measured in blood serum and plasma from 46 cystic fibrosis patients with chronic Pseudomonas aeruginosa colonization before and after treatment. The overall outcome in each patient was evaluated by means of a pondered sum of clinical, chest X-ray and lung function scores. Biochemical markers were related to the overall clinical improvement: haptoglobin, ceruloplasmin, fibronectin and alpha-1 glycoprotein showed a good sensitivity (64-70%), specificity (60-70%) and positive predictive value (86-89%). Granulocyte elastase showed a similar sensitivity (67%) and positive predictive value (85%) but a lower specificity (33%). The negative predictive value was generally poor (32-39%). Our data suggest that the combined measurement of some markers of inflammation and of conventional clinical parameters, may help in evaluating the efficacy of anti-infective treatment in cystic fibrosis.

We studied the prevalence, epidemiological features, symptoms, diagnosis, treatment and outcome of invasive aspergillosis in AIDS patients in Italy.All patients affected by both aspergillosis and AIDS hospitalized between January 1986 and April 1997 (before highly-active antiretroviral therapy, HAART) in four Italian Department of Infectious Disease. Patients were included in the study only if culture, cytology or histology showed firm evidence of Aspergillus infection. Invasive aspergillosis was defined as the presence of characteristic, closely septate hyphae with repeated acute angle branching in either biopsy materials or percutaneous aspirates from tissues other than the lung. Hyphae were identified using hematoxylin-eosin and methenamine silver stain.During the study, 54 out of 2,614 patients admitted with AIDS showed aspergillosis (2.1%). The disease usually occurred in patients with50 CD4 cells/mm(3). Aspergillosis was associated with neutropenia and steroid treatment. Nonspecific symptoms were frequently encountered. Fever and cough were both present in70% of the cases of pulmonary aspergillosis. Biopsy specimens were analyzed for definitive diagnosis. Invasive aspergillosis is usually treated with amphotericin B, but in 90% of the cases this did not prevent death.In AIDS patients with neutropenia and long-term steroid therapy, it is important to consider invasive aspergillosis in the differential diagnosis of opportunistic infections.

The tolerability of and adherence to intermittent short-term rifabutin-isoniazid preventive treatment was assessed in subjects dually infected with Mycobacterium tuberculosis and the human immunodeficiency virus (HIV). In a randomised, open-label, phase II pilot study, 44 subjects received either rifabutin 300 mg and isoniazid 750 mg twice weekly for 3 months (group A, n = 16) or the same regimen with rifabutin at 600 mg (group B, n = 14), or isoniazid 300 mg/day for 6 months (group C, n = 14). Three, two and four subjects in groups A, B, and C, respectively, did not complete their treatment (one case of flu-like syndrome in group B; one methadone withdrawal syndrome in group A; and patient decision in two cases in group A and four in group C). Overall, adverse events were reported by four, nine, and seven subjects in groups A, B and C, respectively. Intermittent combined rifabutin + isoniazid for 3 months had lower default rates than daily standard isoniazid for 6 months. The regimen with rifabutin at 300 mg dose compared favourably to standard isoniazid, and warrants larger efficacy studies to assess its role for the prevention of latent tuberculosis in HIV-infected subjects.

Human neutrophils were isolated both from peripheral blood (PB) and from aseptic inflammatory exudates obtained by the Senn's skin window technique (SW). The respiratory burst (O2- production) induced by in response to n-formyl-methionyl-lencyl-phenylalanine (fMLP) and by serum-treated zymosan (STZ) was investigated using a microplate assay. SW neutrophils were primed to enhanced fMLP-dependent O2- production in response to fMLP but not to STZ. Pentoxifylline, a cAMP-elevating drug, dose-dependently inhibited the respiratory burst in any experimental condition, but the dose-effect curves were markedly different according the stimulant used and the source of the cells. With fMLP as stimulant, a significant inhibition of the O2- production by PB neutrophils was obtained using doses of 10 micrograms/ml, while SW neutrophils were inhibited only by doses equal or higher than 100 micrograms/ml. With STZ as stimulant, the inhibition of the respiratory burst of PB neutrophils and of SW neutrophils was obtained only with doses higher than 400 micrograms/ml and 1 mg/ml respectively. Pentoxifylline dose-dependently (10 micrograms/ml to 1 mg/ml) increased the intracellular adenosine 3'-5'-cyclic monophosphate (cAMP) to the same extent in SW and in PB neutrophils. These data indicate that the priming of neutrophil oxidative metabolism by in vivo inflammation is associated with an increase in the resistance to the regulating effect of cAMP on the fMLP-dependent activation pathway of NADPH oxidase. The fact that therapeutic doses of pentoxifylline do not inhibit the respiratory burst of primed neutrophils may have relevance in the interpretation of the clinical effects of this drug.

To identify the types of medical devices causing needlestick injuries among Italian health care workers, to document the device-specific injury rates and time trends for different hollow-bore needles, and to compare injury rates from these devices with those reported in the United States.Longitudinal survey.Twelve Italian acute care public hospitals.Data were obtained from a multihospital surveillance database on the number of total injuries reported in each device category. Hospitals provided the corresponding number of devices used annually for each needle type.Number of needlestick injuries by type of hollow-bore needle per 100,000 devices used per year.A total of 2524 injuries from hollow-bore needles were reported. Disposable syringes/hypodermic needles accounted for 59.3% of injuries, followed by winged steel needles (33.1%), intravenous catheter stylets (5.4%), and vacuum-tube phlebotomy needles (2.2%). Intravenous catheter stylets had the highest needlestick injury rate (15.7/100,000 devices used), and disposable syringes had the lowest needlestick injury rate (3.8/100,000). In contrast to the other devices, the injury rate from winged steel needles increased from 6.2 per 100,000 in 1990 to 13.9 per 100,000 in 1992.The device-specific needlestick injury rates in Italy are similar to those reported in the United States, suggesting similar exposure experience in two countries. However, in contrast to the United States, needleless intravenous access is standard practice in Italy and thus eliminates one potential risk to Italian health workers. Implementation of safer equipment, such as shielded or retracting needles, and continuing training programs are needed to further reduce the hazards that health care workers face.

In patients with chronic diarrhea associated with acquired immunodeficiency syndrome, bile acid malabsorption, very rarely investigated, may have an important pathogenetic role.In this 1-yr prospective study, 15 patients with AIDS-associated chronic diarrhea and 10 AIDS-controls were studied for bile acid malabsorption by means of the SeHCAT abdominal retention test. The patients with diarrhea underwent the glucose hydrogen breath test to identify any bacterial proliferation in the small bowel.In the chronic diarrhea group, only one case of small bowel bacterial overgrowth, and seven cases (47%) with generally severe bile acid malabsorption, were observed. Among the controls, only one had an abnormal low SeHCAT retention. In both cases with Cryptosporidium infection, the SeHCAT test was pathological.These data are of clinical importance for the pathogenetic study of AIDS-associated chronic diarrhea and for specific treatment with cholestyramine.

Endothelial intercellular adhesion molecule-1 (ICAM-1) and glycoprotein E-selectin (ELAM-1) allow the homing of leukocytes to inflammation sites. A circulating form of ICAM-1 markedly increases in inflammatory CNS disorders. In the present study, the serum levels of ICAM-1, ELAM-1 and tumor necrosis factor-alpha (TNF-alpha) were measured in patients with acute (AIDP) and chronic (CIDP) inflammatory demyelinating polyneuropathies and cryoglobulinemic neuropathy (CGN). Immunoenzymometric assays revealed increased sICAM-1 levels in some of these patients; furthermore, high titres of ELAM-1 and TNF-alpha were detected in two patients with AIDP and one patient with CGN. Our data extend previous observations on inflammatory PNS disorders by showing that, in addition to ICAM-1, ELAM-1 also represents a useful marker of endothelial activation and that, taken together, the two molecules may serve as an indicator of specific pathogenetic mechanisms.

Purified leukemic cells from 30 acute myeloid leukemia (AML) cases at diagnosis were investigated for the presence of interleukin 8 (IL-8) mRNA by Northern blot analysis. IL-8 specific transcripts were detected in uncultured blasts in 14/30 cases, 10/14 from patients with M4-M5 and 4/14 from cases with M0-M3 morphology. The transcript expression was associated with the detection of IL-8 molecule in blast cells by immunostaining performed on cytospin preparations. After 24-hour culture, a strong up-regulation or the appearance in cases negative before culture of IL-8 mRNA was observed in all cases tested, and culture supernatants contained high amounts of IL-8. Our data demonstrate that leukemic cells in AML are equipped with the functional apparatus for IL-8 production. Since IL-8 displays a wide range of biological activities, including the regulation of some membrane molecules relevant to adhesion and migration processes, its production by AML blasts might be of relevance for the pattern of leukemic growth.

Twenty burned patients divided in three groups according to the severity of the lesions were investigated at 1- or 2-day intervals for up to 5 weeks after injury. Plasma elastase levels were elevated during the first day after injury and were correlated with the area of the burns. However, plasma elastase was rapidly bound and inactivated by protease inhibitors. Leucocyte counts, fever and the concentration of alpha-1-proteinase inhibitor were not correlated with the extent of the burn. The rise of plasma elastase was not accompanied by consumption of the elastase inhibitory capacity (EIC) of plasma, which increased to a plateau around day 5. The EIC values were in accord with the rise of alpha-1-proteinase inhibitor, the major anti-elastase agent in plasma. Studies of blister fluid in eight patients showed that the elastase content was higher than that of corresponding plasma, while the concentration of alpha 1-proteinase inhibitor and the EIC were comparable with those of plasma. Measurements of the levels of tumour necrosis factor released by stimulated macrophages in five patients with major burns showed no significant increase compared with controls.

Fibronectin release from cultured macrophages, derived from human blood monocytes, was measured during incubation of the cells with increasing concentrations of vitamin D3 metabolites or of aminobutane bisphosphonate (AHButBP) or dichloromethylene bisphosphonate (Cl2MBP), two powerful inhibitors of bone resorption. The bisphosphonates significantly inhibited fibronectin release at 10(-8) M concentration and this inhibition was almost complete at 10(-5) M concentration. Opposite results were observed when the cells were incubated with vitamin D3 metabolites: the stimulation of fibronectin release was specific for 1,25-dihydroxyvitamin D3 relative to other vitamin D3 metabolites (1,25-dihydroxyvitamin D3 greater than 25-hydroxyvitamin D3 greater than 24,25-dihydroxyvitamin D3).

Serum samples from 66 seropositive subjects (56 with a history of intravenous drug abuse), including asymptomatic carriers and patients with persistent generalised lymphadenopathy (PGL), AIDS related complex (ARC), and AIDS, were tested by indirect immunofluorescence on rat tissue sections and HEp-2 cells for the presence of antibodies to nuclei, smooth muscle, intermediate filaments (anti-IMF) and microfilaments (anti-MF). Counterimmunoelectrophoresis was also used to detect antibodies to extractable nuclear antigens. Smooth muscle antibodies with the V pattern or antinuclear antibodies, mainly of the speckled type, or anti-IMF, occurred in 35 cases, being widely distributed in all groups. Such an autoantibody response resembles the "viral" autoimmunity described in various infectious diseases and in particular that of non-A, non-B post-transfusion hepatitis. Autoantibodies may be of some prognostic relevance, as the prevalence of smooth muscle antibodies V increased as the disease progressed (asymptomatic carriers 20%, those with PGL 29%, those with ARC 47%, and those with AIDS 63%. In the PGL group autoantibody positivity correlated with the presence of skin anergy. The fact that autoantibodies were more frequently detected in patients with circulating immune complexes suggests that these can contain autoantibodies and the corresponding autoantigens.

Thirteen biochemical markers of infection and inflammation were measured during anti-Pseudomonas therapy in cystic fibrosis (CF) patients with respiratory exacerbation. The assessment of some of these markers is thought to be helpful in the evaluation of efficacy of antibiotic therapy in CF.

A clinical epidemiological survey of Legionella pneumophila infections occurring in Italy between 1 December 1985 and 31 May 1986 was carried out to evaluate the incidence of sporadic, epidemic and nosocomial L. pneumophila pneumonia. A total of 355 cases of pneumonia were studied of which 11.5% were due to Gram positive bacteria, 11% were due to Gram negative bacteria, 7.9% were due to Mycoplasma pneumoniae, 4.5% were due to L. pneumophila and 8.5% were due to sundry aetiological agents. The remainder (45.6%) could not be diagnosed accurately. In addition, the anti L. pneumophila antibody titres were assessed. The results are discussed in terms of the occurrence of the disease in Italy and regarding the importance of considering the possibility of legionellosic aetiology when diagnosing pneumonia.

Each albumin variant, its source, and ethnic origin are listed in the Appendix. Many of the new variants were found in several individuals in a single family. In other cases the variants were found in one or more individuals in a single village. The electrophoretic comparisons were made in the three starch-gel systems, acetate-EDTA at pH 5.0, tris-lithium-succinate-citrate at pH 6.0 and tris-EDTA-borate at pH 6.9, used previously (Weitkamp et al. 1973). RESULTS The results of the comparison of 30 new or recently described serum albumin variants with 20 variants previously distinguished using three starch-gel electrophoretic systems are presented in Table 1. Although the amount of separation reported here for the 20 variants previously described differs slightly from the earlier results, due to minor variations in electrophoretic conditions, the conclusions regarding their mobility relative to normal albumin and to each other remain the same. Three new variants, RS I, Xavante, and Yanomama-2, all slowly migrating, have been identified. Electropherograms showing these variants adjacent

Early changes in lipid metabolism and appearance of atherosclerosis risk factors play a key role in the development of cardiovascular disease of chronic renal failure (CRF). In the effort to evaluate the effects of protein restricted diet on dyslipidemia, we studied 122 patients with CRF (S-creatinine 1.3-9 mg/dl); 58.2% of whom were on antihypertensive drugs treatment. Patients had been separated into 6 groups: group 1 was kept on a free diet; groups 2, 3, 4, 5, 6 were kept on a protein-restricted diet from 12, 24, 36, 48, 60 months, respectively. We found hypertriglyceridemia, pathologic levels of esterified cholesterol in high density lipoprotein (HDL-C) and pathologic apolipoprotein A1/B ratio in group 1; the comparison with other groups--whose values were normal range after 12, 24 months of treatment--showed significant differences. The lipidic parameters were independent of the duration of CRF and of patients' age. Serum creatinine showed a significant correlation with tryglicerides and HDL-C values only in group 1. Total cholesterol and apolipoprotein B were significantly greater in hypertensives than in normotensives. In our opinion, a moderate restriction in protein intake could be effective in preventing and in halting the early alterations of lipid metabolism in CRF.

On the basis of personal experience, the microbiological, epidemiological, clinical and therapeutic features of Pneumocystis carinii pneumonia are analysed.

The authors estimated E, E.A.C., and E-active rosette in 13 chronic active hepatitis (C.A.H.), 5 chronic persistent hepatitis and 4 cirrhosis. The results showed a significant decrease of the concentration of peripheral blood T-lymphocytes in patients with C.A.H., C.P.H., cirrhosis. E.A.C. rosette forming cells were not significantly different from the control population in all groups studied. E-active rosette were decrease in patients with C.A.H. and C.P.H. and were increased after stimulation by P.P.D. (5 U.V.I.) in C.A.H. and C.P.H. and in control group studied.

The Authors analyse the results of their tryls about the markers of HBV and HAV acute hepatitis (HBsAg, anti-HAVAb) by R.I.Z. method. HAVAb was in 75% of the cases, its meaning was of post-contact with HAV. The title of HAVAb was effected in 18 patients with viral acute hepatitis; the results were 3 cases of HAV acute hepatitis and in other 7 cases no Ano B viral acute hepatitis. The 58.3% of acute viral hepatitis was HBsAg positive, the study of other markers of HBV and the title of HAVAb showed a viral acute hepatitis caused by HBV. We were not able to demonstrate the viruses which caused 7 HBsAg negative acute virale hepatitis, anti-HBcAg was in 97.8% of HBV acute hepatitis; its the most sensitive of HAV past-infection. The system c-anti-e was in 78.2% of HBsAg viral acute hepatitis. The persistence after 7th week of illness of HBeAg coincided with the hepatitis cronicity. On the contrary anti-HBeAg has not always a protective meaning.

Plasma fibronectin (FN) concentrations were measured by an immunonephelometric method in cord blood from 76 healthy newborns (45 full term and 31 preterm), in 181 healthy children, from 1 month-15 years old, divided by age into seven groups and in 81 control adults. FN levels in newborns were 35% of those of adults. There was no difference between plasma FN in term and preterm (gestational age greater than 30 weeks) infants. Preterm newborns below the gestational age of 30 weeks showed particularly low FN levels. No clear correlation was seen with birth weight. In the child population, a strong increase in FN values was noted in the first 6 months of life, rising to 65% of the normal adult values. No further increase occurred until puberty.

The authors have looked for the markers of HBV by R.I.A. method (HBsAg, anti HBsAg, HBeAg, anti HBeAg, anti HBcAg), of HAV (by measurement on two samples of HAVab or by measurement of HAVab IgM), the immune-complexes (I.C.C.) by C1q solid-phase binding assay method with E.L.I.S.A. with determination after division I.C.C. of HBsAg by R.I.A. method in 33 cases of HBsAg negative acute viral hepatitis. The 9% (3 cases) were HAV acute hepatitis, the 42,4% (14 cases) no A no B acute hepatitis, the 36,3% (12 case) were HBV acute hepatitis, in 9 anti HBcAg positive cases the I.C.C. with HBsAg positive after division resulted positive, the 12,3% (4 cases) had a positivity for HAVab by stereoconversion (2 cases) or HAVab IgM (2 cases) with HBsAg positivity after division I.C.C. This result puts a nosologic problem about the 4 cases of acute viral hepatitis, which, from epidemiological and clinical point of the view are HAV acute hepatitis.

The clinical features of our cases demonstrated some of the already known characteristics of the variable spectrum of HIV infection. DA are the most important risk category in Italy. 10% of the ARC cases evolved into AIDS during a 12-month follow-up, on average. The most frequent OI in our AIDS cases were PCP, C. albicans esophagitis and chronic mucocutaneous ulcers. An high percentage of neurologic involvement from HIV was observed, and malignancies were encountered in AIDS (3 KS and 1 undifferentiated B lymphoma) as well as in ARC (1 Hodgkin's lymphoma). Statistically, significant worsening of the immunologic situation is evident as the disease progresses from LAS to AIDS. Activated B lymphocytes represent most of the cells of the germinal center during the hyperplastic stage of lymphadenopathy. Reversal of the T4/T8 ratio appears early during the initial stage of lymphadenopathy and is due to a decrease of CD4 and a relative increase of CD8. Also, destruction of the follicular dendritic cells is an early feature which becomes more evident as the disease advances and the lymph node evolves toward progressive involution. Activated B-lymphocyte augmentation with polyclonal Ig secretion appears to be related to T-independent B stimulation by coinfection such as CMV, EBV and HBV. The increase of cytotoxic/suppressor lymphocytes seems to be partly related to the excessive activation of B lymphocytes and partially directed to the cells infected by HIV or coated with its proteins (6,7,8,9). The destruction of follicular dendritic cells has been interpreted not only as a killer effect of the virus but also as a result of the intervention of CTL sensitized to the cells containing the virus (10,11). Their destruction may contribute to the impaired recognition of soluble antigen which is one of the main features of the immune deficiency of HIV infection (9,13,16).