A Bruno

Policlinico San Matteo Pavia Fondazione IRCCS, Ticinum, Lombardy, Italy

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Publications (41)74.8 Total impact

  • Journal of Medical Microbiology - J MED MICROBIOL. 01/2010; 59(6).
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    ABSTRACT: Entamoeba histolytica is an intestinal parasite and the causative agent of amoebiasis, which is a significant source of morbidity and mortality in developing countries. Although anti-amoebic drugs such as metronidazole, emetine, chloroquine and nitazoxanide are generally effective, there is always potential for development of drug resistance. In order to find novel targets to control E. histolytica proliferation we cloned, expressed and purified thymidine kinase (Eh-TK) and uridine-cytidine kinase (Eh-UCK) from E. histolytica. Eh-TK phosphorylates thymidine with a Km of 0.27 microm, whereas Eh-UCK phosphorylates uridine and cytidine with Km of 0.74 and 0.22 mM, respectively. For both enzymes, ATP acts as specific phosphate donor. In order to find alternative treatments of E. histolytica infection we tested numerous nucleoside analogues and related compounds as inhibitors and/or substrates of Eh-TK and Eh-UCK, and active compounds against E. histolytica in cell culture. Our results indicate that inhibitors or alternative substrates of the enzymes, although partially reducing protozoan proliferation, are reversible and not likely to become drugs against E. histolytica infections.
    Parasitology 05/2009; 136(6):595-602. · 2.36 Impact Factor
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    ABSTRACT: Malaria is a common, life-threatening infection in endemic tropical areas and one that presents a diagnostic challenge to laboratories in most non-endemic countries. A rapid and accurate diagnosis is a prerequisite for effective treatment, especially for the potentially fatal cases of Plasmodium falciparum infection. In the present, multi-centre study, the performances of a rapid diagnostic test (NOW) Malaria) and several, commercial, PCR-based assays (AMS61, AMS42, AMS43, AMS4 and AMS45) were compared against the results of microscopical examination of bloodsmears (the current 'gold standard'). The subjects were either non-European immigrants (N=135) or international travellers (N=171). There was good concordance between the results of all the detection methods, with kappa values of >0.8. Although the NOW Malaria rapid test was both sensitive (100%) and specific (100%) in detecting P. falciparum infections, it was less specific (93.1%) and sensitive (90.7%) in identifying the other Plasmodium species. The results from the AMS61 assay, designed to detect any malarial infection, generally parallelled those of the microscopy (kappa = 0.89), giving a specificity of 98.2% and a sensitivity of 91.0%. Although the use of species-specific molecular primers to identify pure infections with P. falciparum and P. vivax gave results that were in good agreement with those of the microscopy, the subjects who had apparently pure infections with P. ovale or P. malariae were always found PCR-negative. Compared with the standard microscopy, both the NOW Malaria test and the PCR-based assays were therefore poor at identifying mixed infections. The NOW Malaria test and the PCR-based assays clearly need to be improved, particularly for the correct identification of infections with Plasmodium spp. other than P. falciparum, including mixed infections. For now, expert microscopy must remain the mainstay of the laboratory diagnosis of malaria.
    Annals of Tropical Medicine and Parasitology 05/2007; 101(3):195-204. · 1.31 Impact Factor
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    ABSTRACT: In the present study, 67 patients suspected to be cases of visceral leishmaniasis (VL) were each checked for leishmanial infection by the microscopical evaluation of various biological specimens, in-vitro culture, serology and an assay based on nested PCR. Most (35) of the subjects were immunocompetent (IC) but 32 were immunodeficient (ID) as the result of HIV infection (18 cases), treatment to prevent transplanted organs being rejected (six) or haematological malignancies (eight). Forty-one (61.2%) of the subjects (19 IC subjects, 12 HIV-positive patients, four transplant patients and six patients with malignancies) were considered true cases of VL. For the IC subjects, only the production and microscopical examination of leucocytoconcentrates and cultures of Buffy coats gave sensitivities of <80%, the results of the other methods showing higher sensitivities and almost perfect agreement with the 'gold-standard' diagnoses. For the ID subjects, however, only the serological tests and the PCR gave reasonable sensitivities (of >80%). For the initial diagnosis of leishmaniasis in ID patients, IFAT and western blots may be useful, as, among the present ID patients, they gave sensitivities (of 80.9% and 88.2%, respectively) that were almost as high as that for the PCR, and specificities of 100%. In the diagnosis of VL in either IC or ID patients, the assay based on a nested PCR appeared to be particularly reliable, with sensitivities of 88.9% and 95.2%, respectively, and a specificity of 100% in both groups of patients. The testing of bone-marrow aspirates by PCR revealed very few VL cases who were not found positive when samples of their peripheral blood were checked in the same assay. For both IC and ID subjects therefore, the use of the PCR-based method to test samples of peripheral blood (which can be collected much more easily than bone-marrow aspirates and with much less pain for the subject) is recommended.
    Annals of Tropical Medicine and Parasitology 11/2004; 98(7):667-76. · 1.31 Impact Factor
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    ABSTRACT: The review summarizes the results in the main parasitological topics of our Lab: amoebic infections due to Entamoeba histolytica/Entamoeba dispar complex and to Acanthamoeba spp. respectively, and human infections caused by microsporidia. Different rapid and advanced techniques have been included in the standardized diagnostic protocols for each topic, and a critical comparison among them was made, in order to define the gold standard diagnostic method: a) E. histolytica/E. dispar: in vitro culture, zymodeme typization, biomolecular identification (PCR), immunoenzymatic assay (ELISA) for direct detection in stools of specific surface antigenic lectins; b) Acanthamoeba spp.: in vitro culture, light and ultrastructural characterization, species identification by immunofluorescence method with monoclonal antibodies, in vitro pharmacological studies; c) Microsporidia: ultrastructural (TEM), biomolecular (PCR), biochemical and immunological (SDS-PAGE, Immunoblotting) studies for species identification, use of advanced ultrastructural techniques ("freeze-etching", "deep-etching") in order to deepen the spore wall structure, to study the cytoskeletal function of actin and to define the mode of infection, in vitro pharmacological assays on some inhibitors of chitin-synthases.
    Parassitologia 01/2002; 43 Suppl 1:37-43.
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    ABSTRACT: Technetium 99m tetrofosmin has been introduced as a myocardial perfusion agent, providing similar results to those of thallium 201 and sestamibi in the identification of patients with coronary artery disease. No data are available comparing tetrofosmin and sestamibi imaging in the identification of reversible left ventricular (LV) dysfunction in the same patients. This study compared the results of tetrofosmin, thallium, and sestamibi single photon emission computed tomography at rest in detection of myocardial viability in patients with previous myocardial infarction. Seventeen patients with previous myocardial infarction who were undergoing coronary revascularization were studied. Echocardiography was performed at baseline and 3 months after revascularization to evaluate recovery of LV function. The optimal threshold cutoffs to separate reversible from irreversible dysfunction, as determined by receiver operating characteristic analysis, were 55% of peak activity for both tetrofosmin and sestamibi and 60% for thallium. In all asynergic segments (n = 77) analyzed, tetrofosmin uptake correlated with both sestamibi (r = 0.90, P <.0001) and thallium (r = 0.85, P <.0001) activity. The sensitivity and specificity for reversible dysfunction were, respectively, 70% and 70% for tetrofosmin, 70% and 66% for sestamibi, and 60% and 68% for thallium imaging (all P = not significant). The areas under the receiver operating characteristic curves constructed for tetrofosmin, thallium, and sestamibi activity were 0.74 +/- 0.06 (mean +/- SD), 0.75 +/- 0.06, and 0.74 +/- 0.06, respectively (all P = not significant). Concordance for detecting myocardial viability between tetrofosmin and thallium imaging was found in 67 regions (87%) (kappa = 0.74), and concordance between tetrofosmin and sestamibi imaging was found in 69 regions (90%) (kappa = 0.79). The diagnostic performance of quantitative rest tetrofosmin single photon emission computed tomography in predicting functional recovery after revascularization is comparable to that of both thallium and sestamibi scintigraphy in patients with myocardial infarction and chronic LV dysfunction.
    Journal of Nuclear Cardiology 01/2002; 9(1):33-40. · 2.85 Impact Factor
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    ABSTRACT: Of 550 mentally retarded patients in an Italian institution, 125 (23%) were found to be infected with intestinal parasites. The infections were most frequent in young men, those with severe mental retardation, the chronically institutionalized and those living in older wards. Ninety-four (75.2%) of the parasitised subjects were infected only with protozoa, 25 (20%) only with helminths, and six (4.8%) with protozoa and helminths. Entamoeba histolytica and E. dispar infections were detected, but at low prevalences; in-vitro culture in Robinson's medium and isoenzyme electrophoresis of the cloned amoebic isolates indicated one infection with E. histolytica (zymodeme XII) and two infections with E. dispar (zymodemes I and III). All three Entamoeba-positive subjects were asymptomatic cyst-passers. Antibodies to E. histolytica were detected in seven (1%) of the sera from the 550 patients examined; only one of these was a carrier of an E. dispar strain at the time of investigation. The low prevalences of all the parasitic infections and of the amoebic infections in particular (compared with those observed previously in institutions for the mentally retarded) reflect relatively good facilities and sanitary conditions, an adequate number of well trained staff and good control of the more susceptible subjects.
    Annals of Tropical Medicine and Parasitology 08/2000; 94(5):453-60. · 1.31 Impact Factor
  • Archives of Medical Research 01/2000; 31(4 Suppl):S38-40. · 2.08 Impact Factor
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    ABSTRACT: Four cases of amoebiasis are described: two symptomatic with intestinal and hepatic involvement and two asymptomatic, diagnosed in two, heterosexual, Italian couples. Infection was probably acquired first by the men, via an indirect faccal-oral route, and then transmitted to their partners in the same way. The two amoebic strains isolated, from the woman of one couple and the man of the other, were characterized by electrophoresis as zymodemes II alpha- and XIX of Entamoeba histolytica. These four cases emphasise once more the role of cyst-passers in the spread of infection and the importance of biochemical identification of the amoebic isolates, enabling more specific treatment.
    Annals of Tropical Medicine and Parasitology 01/2000; 93(8):829-34. · 1.31 Impact Factor
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    ABSTRACT: Ileorectal anastomosis (IRA) is a possible surgical treatment for hyperacute and drug-unresponsive forms of ulcerative colitis (UC). UC relapses in the rectal remnant usually are prevented by chronic administration of 5-aminosalicylic acid (5-ASA) in topical formulations. The relationships between intestinal absorption and pattern of luminal spread of 5-ASA enemas are still unknown in patients with IRA. We correlated the absorption of a 5-ASA enema with its spread in the distal bowel of patients with IRA as assessed by 99mTc radioenema imaging. Eight patients with UC in remission and previous IRA received a therapeutic 50-mL 5-ASA enema labeled with 99mTc-sulfer colloid. Absorbed 5-ASA and its major metabolite, acetyl 5-ASA, were measured in plasma, and dynamic images of radiolabeled enema were obtained for 6 h. The retrograde ileal spread (RIS) was determined and expressed as percentage of total enema radioactivity. Plasma levels of 5-ASA and acetyl 5-ASA were measured in six healthy volunteers after administration of the same enema volume with no radiolabeling. The mean 5-ASA plasma level was 0.70 microg/mL (range 0.37-0.95 microg/mL) in patients and 0.96 microg/mL (range 0.78-1.16 microg/mL) in healthy volunteers (P = not significant), and the mean acetyl 5-ASA plasma levels were 0.89 microg/mL (range 0.44-1.19 microg/mL) and 0.84 microg/mL (range 0.51-1.02 microg/mL), respectively (P = not significant). Radioenema imaging allows RIS assessment of patients with IRA. The mean value was 8.5% (range 2%-19.3%) of administered radioactivity, which correlated significantly with the total absorption of 5-ASA in the IRA group (P = 0.033, linear correlation test). Rectal wall contractions recognized by dynamic radioenema imaging were defined as a common cause of RIS episodes. In IRA patients, 5-ASA plasma levels were similar to those in healthy volunteers after administration in enema. Only part of a 50-mL 5-ASA enema reaches the ileum, and radiolabeled imaging shows the degree and number of these RIS episodes. The absorption of 5-ASA can increase in patients compared with healthy volunteers, in the presence of either occasional but significant ileal spread associated with postural factors and abdominal wall contraction or multiple moderate episodes of radioenema backdiffusion related to rectal wall motility.
    Journal of Nuclear Medicine 11/1999; 40(10):1630-6. · 5.77 Impact Factor
  • A Finzi, A Bruno, R Perondi
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    ABSTRACT: Whether tachycardia-dependent paroxysmal AV block, an uncommon complication of exercise stress testing in patients with infranodal conduction disturbances, can result from acute ischemia of the conduction system is still speculative, and is based on post-hoc evidence of right coronary artery disease and abolition of block after coronary angioplasty. In two patients, from a database of 3000 undergoing nuclear exercise stress testing, transient paroxysmal AV block developed 1-4 minutes after the injection of the radionuclide agent. Nuclear perfusion imaging demonstrated stress-induced ischemia of the posteroseptal segments, which corresponds to the anatomical region of the His bundle, and perfusion recovery in the images obtained at rest. Angiography disclosed critical narrowing of the right coronary artery in both cases. Nuclear myocardial perfusion imaging provides noninvasive evidence that transient ischemia of the posteroseptal segment, anatomically corresponding to the His bundle, can result in paroxysmal AV block in patients with severe right coronary artery and chronic infranodal conduction disturbances. The demonstration of the underlying pathophysiological mechanism is useful for selecting the most effective treatment strategy.
    Giornale italiano di cardiologia 11/1999; 29(11):1313-7.
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    ABSTRACT: Cryptosporidium parvum is a coccidian protozoon that causes diarrhoeal enteritis in immunocompetent and immunocompromised humans and other mammals. Sometimes, chiefly in HIV-infected subjects, anatomical sites other than gastro-intestinal tract, such as the biliary and respiratory tree, are involved. We performed an experimental respiratory infection in immunosuppressed albino rats with a C. parvum human-derived isolate, to confirm the possibility of a primary infection at this site and to evaluate the protozoan damages by light and also by transmission electron microscopy (TEM). The animals were infected intratracheally with 1 x 10(6) C. parvum oocysts/ml and, from the 7th day post-infection, biological specimens of trachea, bronchi, lung and ileum were zoopsied. A sole cryptosporidial colonization of the respiratory tract, from the trachea to the median bronchi, without lung parenchyma infection, was observed. Moreover 13/33 (39.4%) rats also developed intestinal infection. TEM study of the respiratory tree specimens demonstrated that cryptosporidia infect either ciliated or goblet cells, and confirmed the role of microvilli in the parasite cell adhesion. The most relevant alterations involved the ciliated cells, with loss of cilia and nuclear and cytoplasmic damages.
    Parasite 10/1999; 6(3):217-22. · 0.82 Impact Factor
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    ABSTRACT: Proteinases play an important role in survival of microorganisms and in pathogenicity of diseases. By using a modified SDS-gelatin-polyacrylamide gel system, proteinases of rat-P.carinii were detected as bands of proteolytic digestion after electrophoresis. P.carinii organisms obtained from dexamethasone immunosuppressed transtracheally infected rats were cultured in spinner flask suspension cultures to minimize host cell contamination. At pH 8.3, seven Pc-specific proteolytic bands were detected in three clusters of different molecular weights clearly different from host cell patterns. By using a range of pH, various preparations of organisms and both infected and uninfected culture media, proteolytic activities have been partially characterized. Elastase secretion has been assessed based on elastin digestion model. Proteinase inhibitors have been tested for their ability to inhibit P.carinii growth in HEL299 short-term monolayer cultures. Results indicate that proteolytic activities are involved in the proliferation of microorganisms since leupeptin exerted in vitro antipneumocystis activity while aprotinin enhanced P.carinii growth.
    Parasite 04/1999; 6(1):9-16. · 0.82 Impact Factor
  • Journal of Nuclear Cardiology 01/1999; 6(1). · 2.85 Impact Factor
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    ABSTRACT: Acanthamoeba keratitis is a severe ocular infection secondary to accidental macro- or microscopic trauma of the cornea. Starting in 1985, a dramatic increase of this infection was recorded along with the spread of contact lens use. This protozoal disease is difficult to treat because of the scarcity of efficacious topical and systemic drugs. We evaluated the in vitro effectiveness of povidone-iodine (PVP-I [Betadine]), an agent with broad antibacterial and antiviral activity, compared to that of chlorhexidine (CXD), a cationic antiseptic, on Acanthamoeba isolates from patients with amebic keratitis. The results showed that PVP-I solution from 0.5 to 2.5% has a better antiamebic activity both on trophic and cystic stages of Acanthamoeba spp. than does CXD.
    Antimicrobial Agents and Chemotherapy 10/1998; 42(9):2232-4. · 4.57 Impact Factor
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    ABSTRACT: An epidemiological survey to characterize Entamoeba histolytica/E dispar isolates from 123 human subjects was carried out in the Wonji area of Central Ethiopia, where an increased incidence of amoebic infection has been reported. In a randomized, coproparasitological study, 93 (75.6%) of the subjects were found positive for at least one species of intestinal parasite: 14 (15.1%) harboured only one species and 79 (84.9%) were infected with at least two species. In-vitro culture in Robinson's medium revealed amoebic parasites in 52 (82.5%) of the 63 cases tested. Of the 29 amoebic isolates successfully stabilized, cloned and characterized by Sargeaunt's electrophoretic technique, 27 (93.1%) were of E. dispar zymodemes (19 of zymodeme I, two each of zymodemes III, V and XI, and one each of zymodemes X and XV) and two (6.9%) were of E. histolytica (zymodeme XIII).
    Annals of Tropical Medicine and Parasitology 04/1998; 92(2):173-9. · 1.31 Impact Factor
  • Parasitology International - PARASITOL INT. 01/1998; 47:203-203.
  • Transplantation Proceedings 01/1998; 29(8):3632-3. · 0.95 Impact Factor
  • Parasitology International - PARASITOL INT. 01/1998; 47:203-203.
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    ABSTRACT: Isoenzyme analysis by starch-gel electrophoresis has proved to be a useful method for the biochemical differentiation of pathogenic Entamoeba histolytica and non-pathogenic E. dispar isolates. Of the known 24 zymodemes, 3 are laboratory-made and have not previously been identified in humans. Parasitology screening was carried out in a psychiatric institution. Two amebic stocks were isolated and characterized that had never previously been found in humans and that have protein patterns identical to that of the laboratory-made zymodeme XX.
    Parasitology Research 02/1997; 83(7):716-8. · 2.85 Impact Factor

Publication Stats

264 Citations
74.80 Total Impact Points

Institutions

  • 1993–2009
    • Policlinico San Matteo Pavia Fondazione IRCCS
      • Dipartimento Malattie Infettive
      Ticinum, Lombardy, Italy
  • 1997–2002
    • Fondazione IRCCS Ca' Granda - Ospedale Maggiore Policlinico
      Milano, Lombardy, Italy
  • 1999
    • University of Milan
      • Istituto di Scienze Dermatologiche
      Milano, Lombardy, Italy
  • 1990–1992
    • University of Pavia
      • Department of Public Health, Neuroscience, Experimental and Forensic Medicine
      Pavia, Lombardy, Italy