Abstract: It is currently unknown if the use of a real-time polymerase chain reaction (PCR) adds value to the diagnosis and follow-up prognosis of patients affected by leishmaniasis. We performed a study using a real-time PCR directed against the alpha-polymerase gene and a semiquantitative PCR that target the SSU ribosomal RNA (rRNA) gene as control for the diagnosis and quantification of parasites in patients with visceral (VL) and cutaneous (CL) leishmaniasis. Our single copy real-time PCR missed one diagnosis of VL compared with the conventional PCR, whereas both PCR methods were able to detect Leishmania parasites in CL. Under anti-leishmania treatment the kinetics of parasitemia were comparable with the two methods. The real-time PCR directed against alpha-polymerase of Leishmania despite being able to make a more accurate quantification of parasites does not add to the decision-making management compared with a semiquantitative PCR, and it is comparatively expensive.
Abstract: Three G3P[9] rotaviruses, detected in children hospitalized with gastroenteritis in Palermo, Italy, were found to be genetically related to strains of either human or feline origin in the VP7, VP4, and VP6 genes. In contrast, in the NSP4 gene the viruses resembled G2P[4] human strains, suggesting a reassortment between AU-1-like and Kun-like strains.
Abstract: OBJECTIVE: The aim of this study was to describe a case of acute acalculous cholecystitis occurring in the course of primary Epstein-Barr virus (EBV) infection. METHODS: The clinical features of the case were analyzed and compared to those of three other similar cases reported in the international literature. RESULTS: All cases occurred in European females with cholestatic hepatitis, presented with gallbladder wall thickening, and recovered uneventfully without the need for surgical intervention. CONCLUSIONS: Acute acalculous cholecystitis may occur during the course of acute EBV infection, especially in patients with cholestatic hepatitis. Clinicians should be aware of the possible involvement of the gallbladder during EBV infection to avoid unnecessary invasive procedures or the overuse of antibiotics.
Abstract: Leishmaniasis is a rarely reported disease among transplant recipients; however, the number of published cases has quadrupled since the beginning of the 1990s. Most cases have been observed in patients living in countries of the Mediterranean basin. Leishmaniasis is most commonly associated with kidney transplantation (77%), and cases are also recorded among patients undergoing liver, heart, lung, pancreas, and bone marrow transplantation. Visceral leishmaniasis (VL) is the most frequently observed clinical presentation, followed by mucosal leishmaniasis and more rarely cutaneous leishmaniasis. Transplant recipients with VL develop the classic clinical form of the disease, which is a febrile hepatosplenic and pancytopenic syndrome. Immunodepression seems to predispose to development of mucosal leishmaniasis caused by viscerotropic strains. Early diagnosis of VL is crucial for patient therapy and outcome; however, this is frequently overlooked or delayed in transplant patients. Pentavalent antimonials are the most commom form of treatment for VL, but have a high incidence of toxicity (34%). Although used in fewer patients, liposomal amphotericin B seems to be better tolerated and should be considered as first-line therapy in transplant recipients.
Abstract: BACKGROUND: The severity of childhood gastroenteritis is generally believed to be age-related rather than aetiology-related. Rotavirus-induced gastroenteritis is more severe than gastroenteritis caused by other enteric pathogens and is also age-related. We thus addressed the question of whether the increased severity of rotavirus-induced gastroenteritis is related to age or to features intrinsic to the agent. STUDY DESIGN: In this multicentre, hospital-based, prospective survey, we evaluated the severity of diarrhoea in rotavirus-positive and rotavirus-negative children up to 4 years of age. Severity was assessed with a score in four groups of age-matched children. RESULTS: Rotavirus was detected in 381 of 911 children. Disease severity was evaluated in 589 cases for which clinical data were complete. The rotavirus-positive and rotavirus-negative groups differed with regards to diarrhoea duration, hospital stay, degree of dehydration and the number of episodes of vomiting. Gastroenteritis was more severe in rotavirus-positive than in rotavirus-negative children. In contrast, none of the main severity parameters differed in the four age groups, irrespective of the presence of rotavirus. CONCLUSIONS: These data provide the evidence that aetiology and not age determines diarrhoeal severity. The demonstration that diarrhoea was more severe in rotavirus-positive children supports the need for a rotavirus vaccine and for studies that address the duration of vaccine protection.
Abstract: BACKGROUND: To overcome some of the limitations of conventional microbiologic techniques, polymerase chain reaction (PCR)-based assays are proposed as useful tools for the diagnosis of visceral leishmaniasis. PATIENTS AND METHODS: A comparative study using conventional microbiologic techniques (i.e., serologic testing, microscopic examination, and culture) and a Leishmania species-specific PCR assay, using peripheral blood and bone marrow aspirate samples as templates, was conducted during an 8-year period. The study cohort consisted of 594 Italian immunocompetent (adult and pediatric) and immunocompromised (adult) patients experiencing febrile syndromes associated with hematologic alterations and/or hepatosplenomegaly. Identification of the infecting protozoa at the species level was directly obtained by PCR of peripheral blood samples, followed by restriction fragment-length polymorphism analysis of the amplified products, and the results were compared with those of isoenzyme typing of Leishmania species strains from patients, which were isolated in vitro. RESULTS: Sixty-eight patients (11.4%) had a confirmed diagnosis of visceral leishmaniasis. Eleven cases were observed in human immunodeficiency virus (HIV)-uninfected adults, 20 cases were observed in HIV-infected adults, and the remaining 37 cases were diagnosed in HIV-uninfected children. In the diagnosis of primary visceral leishmaniasis, the sensitivities of the Leishmania species-specific PCR were 95.7% for bone marrow aspirate samples and 98.5% for peripheral blood samples versus sensitivities of 76.2%, 85.5%, and 90.2% for bone marrow aspirate isolation, serologic testing, and microscopic examination of bone marrow biopsy specimens, respectively. None of 229 healthy blood donors or 25 patients with imported malaria who were used as negative control subjects had PCR results positive for Leishmania species in peripheral blood samples (i.e., specificity of Leishmania species-specific PCR, 100%). PCR and restriction fragment-length polymorphism analysis for Leishmania species identification revealed 100% concordance with isoenzyme typing in the 19 patients for whom the latter data were available. CONCLUSIONS: PCR assay is a highly sensitive and specific tool for the diagnosis of visceral leishmaniasis in both immunocompetent and immunocompromised patients and can be reliably used for rapid parasite identification at the species level.
Abstract: We experienced a case of a 3-year-old boy who presented signs and symptoms of Kawasaki syndrome. Two blood culture sets were processed by the hospital microbiology laboratory using a standard blood culturing system. The anaerobic bottles gave a positive result at day 3 after inoculation. The biochemical profiles produced by the RapID ANA II System showed that the organism was Clostridium baratii with a probability of 99%. Our case highlights the importance of C. baratii as a potential human pathogen and reports the associations with manifestations, which, to our knowledge, have not been previously described concomitantly with a clostridial infection.
Abstract: Twenty-nine staphylococcal isolates from different clinical samples were tested for slime production: phenotypic characterization was carried out using Christensen test (tube test) and Congo red agar plate test (CRA plate test), while the presence and expression of icaA and icaD genes were evaluated by real-time PCR. In 79.3% of studied strains there was a concordance between slime production and presence of icaA and icaD genes, and between lack of slime production and absence of both or only one of the tested genes. In four of five strains where positive phenotype was not associated with the presence of ica genes, gene co-expression (evaluated by mRNA determination) was lacking, while in only a case out of five, there was the presence of transcripts without phenotype. Our study, for the first time, introduces real-time PCR for the detection of both icaA and icaD genes and their mRNA and, furthermore, addresses the relationship between slime phenotype absence and mRNA expression, in clinical isolates of coagulase-negative staphylococci.
Abstract: Infection by an animal-like strain of rotavirus (PA260/97) was diagnosed in a child with gastroenteritis in Palermo, Italy, in 1997. Sequence analysis of VP7, VP4, VP6, and NSP4 genes showed resemblance to a G3P[3] canine strain identified in Italy in 1996. Dogs are a potential source of human viral pathogens.
Abstract: A rotavirus sample collection from 19 consecutive years was used to investigate the heterogeneity and the dynamics of evolution of G1 rotavirus strains in a geographically defined population. Phylogenetic analysis of the VP7 gene sequences of G1P[8] human rotavirus strains showed the circulation of a heterogeneous population comprising three lineages and seven sublineages. Increases in the circulation of G1 rotaviruses were apparently associated with the introduction of novel G1 strains that exhibited multiple amino acid changes in antigenic regions involved in rotavirus neutralization compared to the strains circulating in the previous years. The emergence and/or introduction of G1 antigenic variants might be responsible for the continuous circulation of G1 rotaviruses in the local population, with the various lineages and sublineages appearing, disappearing, or cocirculating in an alternate fashion under the influence of immune-pressure mechanisms. Sequence analysis of VP4-encoding genes of the G1 strains revealed that the older strains were associated with a unique VP4 lineage, while a novel VP4 lineage emerged after 1995. The introduction of human rotavirus vaccines might alter the forces and balances that drive rotavirus evolution and determine the spread of novel strains that are antigenically different from those included in the vaccine formulations. The continuous emergence of VP7-VP4 gene combinations in human rotavirus strains should be taken into consideration when devising vaccination strategies.
Abstract: Mediterranean Spotted Fever is caused by Rickettsia conorii and is transmitted to humans by Rhipicephalus sanguineus, the common dog tick. It is characterized by the symptomatologic triad: fever, exanthema and "tache noire", the typical eschar at the site of the tick bite. In Italy the most affected region is Sicily. The seasonal peak of the disease (from June through September) occurs during maximal activity of immature stage ticks. Severe forms of the disease have been reported in 6% of patients, especially adults with one of the following conditions: diabetes, cardiac disease, chronic alcoholism, glucose-6-phosphate dehydrogenase deficiency, end stage kidney disease. The mortality rate may reach 2.5%. Oral or parenteral administration of tetracyclines or chloramphenicol represent the standard treatment. Recent studies indicate that oral clarithromycin and azithromycin could constitute an acceptable alternative for the treatment of the disease in children; furthermore, they could be recommended during pregnancy.
Abstract: Although the global burden of leptospirosis remains enormous and new aspects of the disease are constantly recognised, little progress has been achieved in the field of leptospirosis therapeutics and queries regarding the utility of antibiotics in the late severe form of the disease remain. From the currently existing data, conclusions on the efficacy of antibiotic administration in severe or late disease cannot easily be drawn, since clinical trials have different selection criteria and may focus on Leptospira serovars with different virulence. However, as a rule the benefit of the doubt should apply. Moreover, new options, such as ceftriaxone, have a superior safety profile to penicillin. In vitro studies have outlined potential antimicrobial candidates such as macrolides and ketolides. Development of a globally accepted subunit vaccine for humans is warranted but is not expected in the near future.
Abstract: Chryseobacterium indologenes is a non-fermentative Gram-negative bacillus that is a rare pathogen in humans. Its occurrence in diabetic children has not been previously reported. In this report, a case is described of C. indologenes bacteraemia possibly associated with the use of a peripheral venous catheter. A 2-year-old boy with type I diabetes mellitus was admitted due to a coma caused by cerebral oedema and was successfully treated for his neurological condition but presented on the tenth day after admission with fever of 40 degrees C, agitation, restlessness, lack of appetite, somnolence and fatigue. His pulse rate was 90 min(-1) and his respiratory rate was 20 min(-1). Laboratory studies revealed a white blood cell count of 4900 mm(-3) with 67% neutrophils and 27% lymphocytes. Two separate blood cultures yielded C. indologenes. Treatment with ceftriaxone was started before the culture results were obtained, and was continued after susceptibility test results were obtained. The patient became afebrile after 48 h, and his general condition improved within 36 h. The infection did not recur. This is believed to be the third case of bacteraemia outside of Asia due to C. indologenes and the first in a diabetic child not otherwise immunocompromised. This case indicates that C. indologenes infection can occur in diabetic children without ventilator or central venous catheter and might be treated with a single agent after in vitro susceptibility tests have been performed.
Abstract: BACKGROUND: Enterococcal meningitis is an uncommon disease usually caused by Enterococcus faecalis and Enterococcus faecium and is associated with a high mortality rate. Enterococcus casseliflavus has been implicated in a wide variety of infections in humans, but never in meningitis. CASE PRESENTATION: A 77-year-old Italian female presented for evaluation of fever, stupor, diarrhea and vomiting of 3 days duration. There was no history of head injury nor of previous surgical procedures. She had been suffering from rheumatoid arthritis for 30 years, for which she was being treated with steroids and methotrexate. On admission, she was febrile, alert but not oriented to time and place. Her neck was stiff, and she had a positive Kernig's sign. The patient's cerebrospinal fluid was opalescent with a glucose concentration of 14 mg/dl, a protein level of 472 mg/dl, and a white cell count of 200/muL with 95% polymorphonuclear leukocytes and 5% lymphocytes. Gram staining of CSF revealed no organisms, culture yielded E. casseliflavus. The patient was successfully treated with meropenem and ampicillin-sulbactam. CONCLUSIONS: E. casseliflavus can be inserted among the etiologic agents of meningitis. Awareness of infection of central nervous system with Enterococcus species that possess an intrinsic vancomycin resistance should be increased.
Abstract: Rotavirus positive samples collected in Palermo, Italy, during 2002-2004 did not react with the G2 type-specific RV5:3 monoclonal antibodies (MAbs) and could be identified as G2 only by RT-PCR genotyping. The genetic variation of VP7 and VP4 antigenic proteins was studied in 14 G2 samples including a selection of both those successfully characterized by serotyping and those failing to be serotyped. The phylogenetic analysis performed on partial VP7 sequences showed a temporal clustering of these strains, with those isolated in Palermo in 2003 belonging to the same lineage of G2 MAbs-unreactive strains identified in UK in 1996-1997 and in Bari, Italy, in 2003-2004. A single amino acid substitution in VP7 antigenic region A, at position 96 (Asp-->Asn), was consistently associated with the loss of antigenic reactivity. Five of the G2 strains were further characterized by sequencing of VP4-encoding genes as belonging to the P[4] type, and separate lineages clustering the strains according to a temporal distribution could be described. VP7 and VP4 antigenic proteins analysis provided evidence that over the last 11 years, at least two different populations of G2P[4] rotavirus strains have been infecting the infant population in Palermo. Considering the role of anti-VP7 and anti-VP4 neutralizing antibodies in rotavirus immunity, the emergence of new VP7-VP4 gene combinations might influence rotavirus circulation in the infant population and should be taken into consideration when devising vaccination strategies.
Abstract: Twenty-two patients with suspected brucellar spondylitis were investigated to evaluate the possible diagnostic role of Sulesomab, a (99m)Tc-antigranulocyte antibody Fab' fragment. Sensitivity and specificity were compared with those of magnetic resonance imaging (MRI). Skeletal involvement was detected by MRI in 11 cases, while leukoscintigraphy indicated normal vertebral uptake in seven of these patients, increased uptake in two patients, and decreased uptake in two patients. Leukoscintigraphy of the 11 patients negative by MRI demonstrated increased uptake in two cases. The sensitivity and specificity of leukoscintigraphy were 27.2% and 81.1%, respectively. Based on these results, leukoscintigraphy is not indicated for the management of patients with suspected brucellar spondylitis.
Abstract: Forty-one children <8 years of age treated for brucellosis with oral minocycline (2.5 mg/kg) twice daily for 3 weeks were recalled and examined to check for dental staining and defects. Dental staining and defects were found in 14 of 41 exposed children (34.1%) and in 30 of 82 matched controls (36.6%), respectively (P > 0.2).
Abstract: OBJECTIVES: To evaluate in a retrospective analysis the efficacy and safety of a 6 day course of liposomal amphotericin B (L-AmB) in infantile cases of Mediterranean visceral leishmaniasis (VL) diagnosed over a 10 year period in Italy. PATIENTS AND METHODS: Patients included were diagnosed as having VL consecutively admitted from December 1992 to December 2001 at four main referral children's hospitals in Italy and treated with six intravenous doses of 3 mg/kg L-AmB given on days 1-5 and 10 (a total dose of 18 mg/kg). Demographic data, nutritional status, underlying diseases, clinical and laboratory findings, and therapy outcome were considered. RESULTS: A total of 164 HIV-negative children (median age 1.6 years; range 4 months to 14 years) were enrolled. All patients were initially cured by the given treatment, and did not present adverse events related to drug infusion. Seven patients (4.3%) had a clinical and parasitological relapse 3-15 months after therapy. All relapses were successfully retreated with 3 mg/kg L-AmB for 10 consecutive days (a total dose of 30 mg/kg). CONCLUSIONS: This study highlights the efficacy (>95%) and safety of the six dose L-AmB regimen and validates it as a first-line treatment for Mediterranean VL in children.
Abstract: Among the known human rotavirus serotypes, types G1-G4 are ubiquitous and account for >80% of global human rotavirus strains. Since 1994 an increase in reports of G9 serotype isolates has been observed in both developed and developing countries. In the winter season of 1999-2000 we detected the appearance of G9 rotavirus in Palermo, Italy, accounting for 19% of all rotaviruses circulating in our geographical area. Some of these Italian G9 isolates have been submitted to partial sequencing of their VP7 gene. All of them showed complete nucleotide identity suggesting a clonal origin. The Italian VP7 sequences were found to be very closely related to that of other G9 strains recently isolated in Europe, America, Africa and Asia. Our results confirm that G9 strains circulating worldwide since 1994 are closely related genetically in their VP7 genes.
Abstract: Abstract A 1-year study involving 157 gastroenteritis samples was conducted to investigate the role of human astrovirus (HAstV) as a cause of gastroenteritis in Italian children aged < 2 years. The overall incidence of HAstV was 3.1%. Most cases occurred between March and May, and four of the five isolates were of the HAstV-1 type, the other being HAstV-3. Analysis of genetic variability showed that the three HAstV-1 isolates collected in 2000 clustered together, but separately from the 1999 isolate. The results indicated that HAstV should be considered as a potential diarrhoeal pathogen in Italian children.
Abstract: Visceral leishmaniasis (VL) is endemic in areas bordering the Mediterranean Sea (Spain, Italy, France, Greece, Morocco, Tunisia) where it is caused by Leishmania infantum and it is transmitted by the bite of hematophagous sandfly belonging to Phlebotomus spp.; dog constitutes the main reservoir of the infection. In comparison with the past, when VL was typically observed more frequently in children, the current ratio of childhood to adult cases is approximately 1:1. The onset of the disease is characterized by a non-specific initial symptomatology; fever, pallor and splenomegaly are always present. Pancytopenia is present very often; the laboratory diagnosis is established by serological tests (indirect fluorescent-antibody assay, immunoassay test, indirect hemagglutination assay) and by demonstration of Leishmania parasites by microscopy, culture or polymerase chain reaction (PCR) in the bone marrow aspirates. The use of PCR performed on peripheral blood has been reported to be highly sensitive for the diagnosis and the follow up of children with VL. Pentavalent antimonial drugs have been used for many decades as standard treatment for VL; in Italy liposomal amphotericin B (AmBisome) is nowadays considered the first-line treatment for VL.
Abstract: In order to evaluate the efficacy and tolerability of a high intravenous dose of rifampin plus oral minocycline (administered daily for 3 weeks) for the treatment of acute brucellosis, we retrospectively reviewed the outcome of 239 consecutive patients (135 adults and 104 children) diagnosed and treated over a 17-year period in Italy. The combination used resulted in 100% response and a relapse rate lower than 2%. Fifty-two (30 adults and 22 children) (29.8%) complained of mild adverse effects including an increase in aspartate aminotransferase (>250 IU) observed in 12 cases and considered related to rifampin and in 11 cases a reversible hyperpigmentation of the tongue attributed to minocycline. A randomized prospective comparative study should be performed to confirm our encouraging results.
Abstract: We report a significant high incidence of infection with G9P[8] rotavirus in Italian children during the winter epidemic season 1999-2000. The study was carried out on 439 children < 4 years hospitalized with acute diarrhea in Palermo. G9P[8] strains constituted 19% of all rotavirus identified and were not associated with more severe forms of gastroenteritis.
Abstract: The clinical and epidemiological characteristics of 111 consecutive cases of visceral leishmaniasis identified from 1980 to 2000 in a Sicilian pediatric hospital were analyzed retrospectively. The mean age of the patients was 1.7 years. All children were HIV negative, but 15% were severely malnourished. Fever and splenomegaly were present in all cases and hepatomegaly in 101 (90.1%) cases. Thrombocytopenia and anemia were both observed in 78 (70.2%) cases and leukopenia in 47 (42.3%) cases. A bone marrow aspirate was obtained in all cases; Leishmania amastigotes were detected in 89 (80.2%) cases. Initial treatment consisted of meglumine antimoniate in 99 (89.2%) patients and amphotericin B in 12 (10.8%) patients. Only two children treated with meglumine antimoniate relapsed. The findings highlight the differences between the cases of visceral leishmaniasis observed in the Mediterranean basin and those observed in other regions. The use of the term "Mediterranean visceral leishmaniasis", rather than the term "kala-azar", is proposed for cases observed in the Mediterranean area.
Abstract: OBJECTIVE: To assess the usefulness of a polymerase chain reaction (PCR) assay amplifying the small subunit rRNA coding region of Leishmania species performed on peripheral blood (PB) and bone marrow (BM) aspirates for the diagnosis and follow-up of visceral leishmaniasis (VL) in children living in the Mediterranean basin. DESIGN: A prospective study was conducted on children consecutively hospitalized over a 1-year period at our Infectious Diseases Department in Sicily (Italy) presenting with fever, hepatosplenomegaly, and/or pancytopenia and a positive Leishmania serology (> or =1:40). RESULTS: Among the 14 patients hospitalized with signs and symptoms suggestive of the disease and a positive serology, we identified 10 cases of Mediterranean VL. PCR performed on PB and BM aspirates was positive in all cases and concordant with microscopy and/or culture performed on BM. Leishmania DNA was cleared from PB a median of 6 days after the start of treatment; during follow-up (median: 9 months; range: 6-12 months) 1 child relapsed. In this case, BM PCR remained positive with rapid reappearance of a positive signal also in PB. CONCLUSIONS: PB PCR allows a rapid and noninvasive parasitologic diagnosis of Mediterranean VL among immunocompetent children and is at least as sensitive as a diagnosis made on the basis of BM aspirates. The lack of disappearance from BM and the reappearance of positive PCR on PB is predictive of clinical relapse. Qualitative and semiquantitative PCR may be the standard method for monitoring response to therapy in immunocompetent children.
Abstract: We conducted an open-label randomized controlled trial to compare the efficacy and safety of clarithromycin (15/mg/kg/day in 2 divided doses for 7 days) with those of azithromycin (10 mg/kg/day in 1 dose for 3 days) in the treatment of children with Mediterranean spotted fever. Until now, there has not been a gold-standard therapy for this rickettsial disease in children. Eighty-seven children were randomized to receive 1 of the 2 drugs. The mean time to defervescence (+/- standard deviation) was 46.2+/-36.4 h in the clarithromycin group and 39.3+/-31.3 h in the azithromycin group. These differences were not statistically significant and both drugs were equally well-tolerated. Clarithromycin and azithromycin could be acceptable therapeutic alternatives to chloramphenicol and tetracyclines for children aged < or =8 years with Mediterranean spotted fever. Azithromycin, because it has a long half-life, offers the advantages of administration in a single daily dose and a shorter duration of therapy, which could increase compliance in children.
Abstract: Till now there is not a gold standard therapy for Mediterranean spotted fever (MSF) in children. Standard treatment for MSF is the administration of tetracycline or chloramphenicol, however both these drugs can cause significant adverse effects in children (tetracyclines can cause staining of teeth, chloramphenicol severe hematological adverse events such as aplastic anemia, gray baby syndrome and hemolytic anemia in patients with the Mediterranean form of G6PD deficiency). We conducted two randomized clinical trials; the first compared clarithromycin versus chloramphenicol: mean time to defervescence was 36.7 +/- 18.1 h in the clarithromycin group and 47.1+/- 21.9 h in the chloramphenicol group (P= 0.047). The second trial compared clarithromycin versus azithromycin and did not show any statistically significant difference: mean time to defervescence was 46.2 +/- 36.4 h in the clarithromycin group and 39.3 +/- 31.3 h in the azithromycin group (P= 0.34). On the basis of these studies we think that clarithromycin and azithromycin could constitute an acceptable alternative to chloramphenicol and to tetracyclines for the treatment of MSF in children
Abstract: The aim of our study was to determine whether the severity of rotavirus gastroenteritis may be related to the different characteristics of infecting viral strains. The severity of clinical symptoms in 401 children with acute rotavirus gastroenteritis was assessed using a scoring system for frequency and duration of vomiting, diarrhea, and fever, as well as the patients' requirements for intravenous rehydration. Rotavirus strains were characterized by determining the electropherotype of their double-stranded RNA, the G type and subgroup by a panel of monoclonal antibodies, and the P type by reverse transcription-polymerase chain reaction. Strains with a short electropherotype, G2P[4] type, and subgroup I were associated with more-severe gastroenteritis and affected children older than those infected with strains with a long electropherotype, G1P[8] or G4P[8] type, and subgroup II. Minor differences in clinical symptoms were also detected in children infected with different long electropherotypes and with G1P[8] and G4P[8] specificities.
Abstract: The aim of this study was to compare pertussis-specific humoral and cellular immunity in children 5 years after a primary vaccination with a combined diphtheria, tetanus, tricomponent acellular pertussis, and hepatitis B vaccine (DTaP-HBV; InfanrixHepB; SmithKline Beecham) with immunity after natural infection. The subjects were 38 children aged 5 to 6 years who received DTaP-HBV at 3, 5, and 11 months of life and 21 subjects of similar ages and sex who acquired pertussis in the first year of life. Immunoglobulin G (IgG) antibody titers against Bordetella pertussis antigens, peripheral blood mononuclear cell-specific proliferation, and the secretion of cytokines were evaluated. After 5 years, only a small proportion of vaccinated and infected children had significant specific concentrations of IgG in serum against all three B. pertussis antigens, and T-cell responses persisted in a minority of subjects. A preferential type 1 cytokine response with the secretion of gamma interferon was observed in the pertussis group, whereas a type 2 skewed response was observed in the vaccinated children; however, the quantitative differences in the cytokines produced by DTaP-HBV and natural infection were minimal. In conclusion, our results show that the immune responses induced by primary pertussis vaccination are qualitatively and quantitatively similar to those seen in children who recovered from natural infection and highlight the need for booster immunization with pertussis vaccines in order to maintain adequate levels of a specific immune response to B. pertussis.
Abstract: Fifty-one children with Mediterranean spotted fever (MSF) were randomized to receive either clarithromycin, 15 mg/kg/day orally in 2 divided doses, or chloramphenicol, 50 mg/kg/day orally in 4 divided doses, for 7 days. Mean time to defervescence was 36.7 h in the clarithromycin group and 47.1 h in the chloramphenicol group (P=.047). Clarithromycin could be an acceptable therapeutic alternative to chloramphenicol and to tetracyclines for children aged <8 years with MSF.
Abstract: BACKGROUND: Compulsory vaccination of children against hepatitis B virus (HBV) infection was introduced in Italy in 1991. PATIENTS AND METHODS: To evaluate the current importance of pediatric HBV infection, we studied 359 HBsAg-positive children admitted to 16 centers in Italy from 1991 to 1998. 185 patients were natives of Italy and 174 (39 immigrants and 135 adopted) came from highly endemic countries (eastern Europe: 60.9%, Asia: 16.7%, Africa: 14.9% and Central and South America: 5.7%). RESULTS: Transaminase Levels were moderately altered in both Italian (mean 134 UI/L) and foreign children (mean 168 UI/L). In total, 77% of ItaLian children and 88% of foreign children tested HBeAg positive. High transaminase levels and HBeAg positivity were more frequent in adopted children. Follow-up of 317 patients showed that the incidence of HBeAg/anti-HBe serum conversion was similar in all cohorts, but in adopted children it occurred at an earlier age and was associated with HBsAg clearance in 5%. CONCLUSION: HBV is not frequent in Italian children today, but it is common among children coming from highly endemic areas. The vaccination of nonimmune native populations must be strongly recommended.
Abstract: The aim of our study was to diagnose and to control three aspects of the evolution of lung disease in CF: the absence of infection, the intermittent colonization and chronic infection by Pseudomonas aeruginosa. Therefore a study of anti-pseudomonas antibodies (Ab) (anti-protease, anti-elastin and antihexo-toxin A) for diagnosis and follow-up of CF patients was considered. Moreover, we related the presence of Ab to the sputum culture, to FEV1, to patient age and to genotype. Tbe Ab were dosed in 121 patients by quantitative ELISA method. Values < 1: 500 were considered negative, values> 1: 500 and < 1:1250 borderline, and > 1:1250 positive. 16.5% of patients did not have Ab, 17% had borderline values and 69.5% had positive values. All the patients with negative Ab had negative sputum culture; 47% of patients with borderline values had at least one positive culture while 53% were negative. 87% of patients with positive values had chronic colonization, 13% intermittent colonization. The increase in the Ab rate is statistically related to a more severe lung disease (p < 0.013). The presence of a severe mutation (?F 508) is related to positive values of Ab. Evaluation of anti-Pseudomonas aeruginosa is an important tool for diagnosis and follow-up of CF lung disease
Abstract: A seminested PCR typing assay has been extended to identify rotavirus strains with the P[14] genotype. The specificity of the method was confirmed by Southern hybridization and by restriction analysis with the enzyme AluI. One out of four human rotavirus (HRV) strains with unusual subgroup-electropherotype linkage but none out of 50 HRV strains with usual linkage was typed as P[14].
Abstract: The spectrum of signs and symptoms of 645 consecutive children diagnosed from 1984 to 1996 with boutonneuse fever (BF), a mild rickettsial disease caused by Rickettsia conorii endemic in the Mediterranean basin, are reported. The major clinical features were fever (97.2%), exanthema (96.1%) and "tache noire" (71.8%). The large series examined permitted the authors to observe some rare or disregarded clinical features of the disease: cases with papulovesicular exanthema, reported previously only in adults who had been infected by R. conorii in Africa; and cases in which the only symptom was an isolated lymphadenopathy. CONCLUSION: R. conorii infection should be considered in patients with lymphadenopathy who live in or have travelled to an endemic area, even when other, more specific features, are not present. Also pox-like vesicular lesions may be caused by this organism, awaiting confirmation by using culture techniques instead of serology. The serological confirmation of BF by immuno fluorescent antibody test is possible only late in the illness.
Abstract: Visceral leishmaniasis (VL) is endemic in Sicily. Although it is a notifiable disease, there is evidence that the actual number of cases is higher than that reported. In 1987, a regional reference center for active surveillance of VL was established and it recorded a total of 284 cases through 1995, a mean of 31.5 cases/year and about four-fold more than previously reported. Of the 284 cases, 150 (53%) were children (< or = 14 years of age), and of the 134 adults, 39 (29%) were coinfected with human immunodeficiency virus (HIV). The commonest viscerotropic zymodeme of Leishmania infantum, MON 1, was identified in 40 (93%) of 43 HIV-negative and eight (57%) of 14 HIV-positive patients. Among 280 patients evaluated (i.e., all HIV-negative and 35 of 39 HIV-positive subjects), 254 (91%) were treated with meglumine antimoniate alone or in combination with other drugs; 23 (8%) received allopurinol or amphotericin B, either conventional or in liposomal form; and three terminally ill patients were not treated. Among the 245 HIV-negative patients, 236 (96%) were successfully cured, while nine (4%) (seven adults) died during the course of antimonial treatment. None of the 35 HIV-positive patients was definitively cured, although mortality was apparently associated with other opportunistic infections.
Abstract: The plasma levels of tumor necrosis factor-alpha and interferon-gamma were measured in 53 consecutive children with serologically confirmed Mediterranean spotted fever and were found to be increased during the acute phase compared with the convalescent phase (tumor necrosis factor-alpha mean 32.17 vs. 4.12 pg/ml, P < 0.0001; interferon-gamma mean 84.17 vs. 2.65 pg/ml, P = 0.0006). Plasma levels of both cytokines were higher in patients with a typical exanthema rather than those with a very mild or no exanthema; tumor necrosis factor-alpha levels were significantly lower in the latter (tumor necrosis factor-alpha 32.17 vs. 9.85 pg/ml, P < 0.0001; interferon-gamma 84.17 vs. 38.14 pg/ml, P = 0.35). Tumor necrosis factor-alpha and interferon-gamma may be harmful or beneficial to the infected host, depending upon the amounts produced and whether they are circulating or confined locally to the site of inflammation.
Abstract: We used liposomal amphotericin B as first-choice treatment of visceral leishmaniasis in 106 immunocompetent children who acquired the infection in a temperate region of southern Europe (Italy) where Leishmania infantum visceral leishmaniasis is endemic. The aim of the study was to identify the minimum total dose of liposomal amphotericin B needed to cure the infection in children and reduce the period of hospitalization. We conclude that the optimal regimen in immunocompetent children with L. infantum visceral leishmaniasis to be a total dose of 18 mg/kg of liposomal amphotericin B (3 mg/kg per day for 5 days, followed by 3 mg/kg administered as an outpatient regimen on day 10).
Abstract: 108 rotavirus strains obtained from children with diarrhea hospitalized in Palermo, Italy, in the years 1990-1994, were examined by seminested PCR to study the relative frequency and distribution of the four most common alleles of the gene 4. Such strains were selected from 344 human rotavirus strains recovered in palermo during those years after characterization by electropherotyping, subgrouping and G serotyping. One hundred and seven of the 108 strains could be classified into P types, the P[8], G1 (38.3%) and the P[8], G4 (52.3%) types being predominant. The unique strain whose P genotype could not be identified showed an unusual combination of long migration electrophoretic pattern and subgroup I specificity.
Abstract: Polyacrylamide gel electrophoresis of nucleic acid extracted from stool samples of diarrhoeic children revealed in 3 out of 690 (0.43%) specimens two electrophoretic bands with a migration pattern characteristic of picobirnavirus ds-RNA. In none of the 92 control children were similar bands detected. No other potential enteric pathogens were found in the patients with picobirnavirus infection.
Abstract: The rapidity and efficacy of a short course of liposomal amphotericin B was evaluated in 29 children affected by visceral leishmaniasis (Leishmania infantum). Their overall health status was assessed using the prognostic inflammatory and nutritional index (PINI), and their haematological status by the reticulocyte count and haemoglobin blood levels. All these quantities were measured on day 0, and 3 and 10 d after starting therapy. A significant decrease of inflammatory signs, associated with an improved reticulocyte count, was recorded after 3 d of therapy. A significant increase of haemoglobin levels was also observed 10 d after the start of treatment. The early reduction of inflammatory signs and the improvement of bone marrow function in most patients confirmed the validity of amphotericin B therapy. The PINI score is helpful in assessing the severity of visceral leishmaniasis and the follow-up of its treatment.
Abstract: To examine the role of enteric adenoviruses (Ad40 and Ad41) in children with acute gastroenteritis, we evaluated 273 children with diarrhoea and 137 without enteric symptoms in Palermo, Italy, during an 8-month period. Stools were tested by two home-made monoclonal-based ELISAs to detected genus-specific adenovirus antigen and to type Ad40 and Ad41. Twenty-five samples (6.1%) were found to contain adenovirus, 18 of which were grown in Graham 293 and in HEp-2 cells. Ad40 and Ad41 were detected in 2.6% of children with diarrhoea and in none in the control group, while non-enteric adenoviruses were obtained from both patients (3.2%) and controls (6.5%). Samples containing Ad40 and Ad41 were positive by the virus isolation procedure in Graham and in HEp-2 cells, showing no distinct growth pattern in these cell lines. The evaluation of a latex agglutination test (Adenolex) and of a commercial ELISA (Adenoclone), respectively available for the detection of genus adenovirus antigen and for the typing of Ad40 and Ad41 suggests that both tests enable the identification of enteric adenoviruses in stool specimens, giving results comparable to our ELISAs.
Abstract: We evaluated liposomal amphotericin B (AmBisome; Vestar, San Dimas, CA) administered to 88 immunocompetent patients (56 children) with visceral leishmaniasis (VL) caused by Leishmania infantum. Thirteen patients received 4 mg/kg on days 1-5 and 10 (total dose, 24 mg/kg), and all were cured; 42 received 3 mg/kg on days 1-5 and 10 (18 mg/kg), and 41 were cured; 32 received 3 mg/kg on days 1-4 and 10 (15 mg/kg), and 29 were cured (amastigotes were not cleared from 1 child, and 2 relapsed). One adult was cured with a total dose of 12mg/kg. The four children who were not cured received 3 mg/kg for 10 days; none had further relapses. There were no significant adverse events. For VL due to L. infantum, we recommended a total dose of AmBisome of > or = 20 mg/kg, given in > or = 5 doses of 3-4 mg/kg over > or = 10 days.
Abstract: During a measles outbreak, 112 serum specimens from 88 hospitalized patients were received in our laboratory for investigation of a morbilliform rash. These specimens (88 acute- and 24 convalescent-phase) were tested for the presence of measles-specific IgM antibodies by a capture EIA (enzyme immunoassay) using peroxidase-conjugated measles virus antigens and by an indirect EIA. Commercially available indirect EIA kits for measles-specific IgM antibodies were also used and compared with our homemade EIAs. Specificity studies included a collection of serum specimens containing rheumatoid factor, antinuclear antibodies or IgM antibodies specific to other viruses, and sera from blood donors and healthy children. Sensitivity of capture EIA and indirect EIA to detect measles IgM was 91.8 and 90.3%, respectively, and specificity was 98.2% for both tests. Specific IgM antibodies were detected in 70.5% of serum specimens at the first day after rash onset and were present for a month following the rash. Among the commercial measles IgM detection assays, EIA "Behring" was found to be a valid alternative for detection of measles virus-specific IgM.
Abstract: Thirty-one patients with visceral leishmaniasis (VL) caused by Leishmania infantum received liposomal amphotericin B (AmBisome) in a multi-centre study. Ten immunocompetent patients (six children) received 1-1.38 mg/kg/day for 21 days, and ten (nine children) received 3 mg/kg/day for 10 days. All were cured without significant adverse events and without relapse during 12-24 months of follow-up. Eleven immunocompromised adults, including seven co-infected with HIV (four with AIDS) received 100 mg (1.38-1.85 mg/kg) daily for 21 days. All were initially considered cured, but eight relapsed clinically and parasitologically at 3-22 months. Liposomal amphotericin B is a new, safe and effective drug for the treatment of VL.
Abstract: 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.
Abstract: To determine whether a nontoxic derivative of pertussis toxin obtained by recombinant DNA technology, PT-9K/129G, is a good candidate for a new pertussis vaccine, we examined the safety and the immunogenicity in children of a vaccine containing 15 micrograms of PT-9K/129G protein and 0.5 mg of aluminum hydroxide per dose. Fifty-three children 12 to 24 months of age and 21 infants aged 2 to 4 months were injected with two and three doses, respectively. The vaccine did not induce significant local or systemic reactions and elicited an increase of antibody titer in more than 98% of the children. The geometric mean of the toxin-neutralizing titers increased after each dose and was 85 units in children given two doses and 196 units in those given three doses. Two children who had detectable antibody levels before the first immunization had a high response (greater than 320 units) to the first vaccine dose. The findings suggest that PT-9K/129G is a promising antigen to be included in the development of acellular pertussis vaccines.
Abstract: In 1988 in Palermo, Italy, the prevalence of antibodies to hepatitis A virus (anti-HAV) in a sample of 490 children 6-13 years old was 10.6%; it increased from 6.3% among children 6-10 years old to 14.7% in children 11-13 years old (P less than 0.01). Compared with findings from a survey conducted in 1978 in the same area, the results of the present study show a significant (P less than 0.01) reduction in the anti-HAV prevalence in both age groups. Anti-HAV prevalence was inversely related to the father's years of education and positively related to the family size. Children of fathers with less than 6 years of schooling had a 3.2-fold risk (C.I. 95% = 1.3-8.1), and children with five or more members in their households had a 2.7-fold risk (C.I. 95% = 1.1-6.4) of previous exposure to hepatitis A virus (HAV) infection. Our findings indicate that exposure of children in Palermo to HAV is decreasing significantly, probably because of improvements in socio-economic conditions during recent years; however socio-demographic factors appear to be important determinants of infection.
Abstract: During 1985-89, an epidemiological survey was conducted in Palermo, Sicily (Southern Italy) on group A human rotavirus (HRV) strains which cause gastroenteritis in infants and young children. Two hundred and thirty eight HRV strains were characterized for subgroup and serotype using monoclonal-antibody-based ELISA systems, and for electropherotype using polyacrylamide gel electrophoresis. Subgroup II strains were largely predominant, constituting 218/238 of the positive stool samples (91.6%). Among the serotypes, 192/238 strains (80.7%) were serotype 1 and 16 strains (6.7%) were serotype 4; serotype 2 circulated intermittently and serotype 3 was nearly absent (only one subgroup I strain was detected). Two electropherotypes, bbba and cbba, accounted for the largest proportion of the 345 HRV strains examined, 74 (21.4%) and 222 (64.3%) strains, respectively. Unexpected combinations of subgroup, serotype and electropherotype were detected in 5 subgroup I strains, of which 4 possessed a "long" RNA pattern (1 serotype 3 and 3 serotype 4 strains) and one a "short" RNA pattern (a serotype 4 strain). In addition, 4 group C HRV strains (atypical HRV or pararotaviruses) were detected on the basis of electropherotype. These findings emphasize the need for continuous surveillance of HRV infections in different geographic areas of the world in order to detect the appearance of new strains early and to adopt adequate strategies for vaccine preparation and administration.
Abstract: The prevalence of IgG antibodies to Bordetella pertussis in a sample of 615 1-12-year-old unvaccinated children in Palermo was estimated by ELISA. The overall prevalence was 56%; it increased from 24% in one to three-year-old children to 67% in 11-12-year-old children (p less than 0.01). IgG antibody prevalence was not associated with the father's years of schooling (OR 1), nor with the family size (OR 1.3; C.I. 95% = 0.8-2.2). For children aged one the three years, serological results showed that the history of pertussis reported by parents in questionnaires gave high specificity (93.2%) and negative predictive value (85.4%). Our seroepidemiological study evidences a great exposure of children to B. pertussis in Palermo, with a high proportion of infections occurring after three years of age.
Abstract: An ELISA assay was performed to detect antigens of Brucella melitensis directly in the blood of patients affected by Brucellosis. Disposable polystyrene microtiter plates were coated with rabbit immunoglobulins anti-Brucella melitensis antigens and then incubated with sera of Brucellosis patients and sera of not infected normal subjects as a control, to standardize the conditions of the different steps of the assay. The level of the blood-containing Brucella antigen bound to the plate was measured by addition of anti-Brucella melitensis antiserum conjugated with alkaline phosphatase followed by incubation with the specific enzyme substrate. Sera from 9 Brucellosis patients not undergoing therapy were tested by this ELISA assay, and all showed values significantly higher than the control. A pool of 96 sera from normal subjects not infected with Brucella melitensis was used as a negative control. In addition, a different group of 8 sera from patients with Brucellosis undergoing therapy were also analyzed, but no difference in the ELISA value was observed between the two groups with or without therapy. The ELISA assay described in this paper could be a reproducible, sensitive and suitable test to detect Brucella-antigens in the blood of Brucellosis patients and it could be used in addition to the more common methods for a more thorough diagnosis of Brucellosis.
