Serological study on Chlamydophila pneumoniae in patients with community-acquired pneumonia.

This study aimed to evaluate the incidence of Chlamydophila pneumoniae antibodies in patients with community acquired pneumonia (CAP) by a new ELISA test (EIA CP-IgG, IgA, IgM--Eurospital, Trieste, Italy). From January 1999 to July 2001 141 patients with clinical signs of CAP were enrolled in sixteen Italian Hospitals. Specific IgM and IgG antibodies anti-C. pneumoniae in serum and IgA in both serum and sputum were detected. At a primary inspection (time T-0) serum and sputum samples were taken from 115/141 patients, whereas serum was collected from only 100/141 patients after 30 days (time T-30). At T-0 24/115 (20.8%) patients showed serological markers thus suggesting an acute C. pneumoniae infection. In 23/24 patients the overall serological pattern found at T-0 was confirmed at T-30. In 32/115 patients (27.8%) serological markers of C. pneumoniae past infection were found positive and were confirmed 30 days later. These data support the role of C. pneumoniae as an important aetiological agent of CAP throughout different geographic areas of Italy. The test was suitable for the laboratory diagnosis of C. pneumoniae infection. In particular, the presence of specific IgA anti- C. pneumoniae in both serum and sputum proved useful to define different stages and evolution of infection.

Burden of unidentifiable mycobacteria in a reference laboratory.

Modern identification techniques at the genomic level have greatly improved the taxonomic knowledge of mycobacteria. In adjunct to nucleic acid sequences, mycobacterial identification has been endorsed by investigation of the lipidic patterns of unique mycolic acids in such organisms. In the present investigation, the routine use of high-performance liquid chromatography (HPLC) of mycolic acids, followed by the sequencing of the 16S rRNA, allowed us to select 72 mycobacterial strains, out of 1,035 screened, that do not belong to any of the officially recognized mycobacterial species. Most strains (i.e., 47) were isolated from humans, 13 were from the environment, 3 were from animals, and 9 were from unknown sources. The majority of human isolates were grown from the respiratory tract and were therefore most likely not clinically significant. Some, however, were isolated from sterile sites (blood, pleural biopsy, central venous catheter, or pus). Many isolates, including several clusters of two or more strains, mostly slow growers and scotochromogenic, presented unique genetic and lipidic features. We hope the data reported here, including the results of major conventional identification tests, the HPLC profiles of strains isolated several times, and the whole sequences of the 16S rRNA hypervariable regions of all 72 mycobacteria, may encourage reporting of new cases. The taxonomy of the genus Mycobacterium is, in our opinion, still far from being fully elucidated, and the reporting of unusual strains provides the best background for the recognition of new species. Our report also shows the usefulness of the integration of novel technology to routine diagnosis, especially in cases involving slow-growing microorganisms such as mycobacteria.

Invasive aspergillosis in haematological malignancies: clinical findings and management for intensive chemotherapy completion.

Sixty-one cases of Aspergillus infection (35 acute myeloid leukemia, 15 acute lymphoid leukemia, one myelodysplastic syndrome, two aplastic anemia, eight non-Hodgkin's lymphoma) seen in our department between January 1989 and July 1999 were studied retrospectively to evaluate the clinical characteristics, to ascertain the factors that influenced the outcome from mycotic infections, and whether early diagnosis and prolonged therapy permitted completion of scheduled intensive chemotherapy and bone marrow transplantation (BMT) without fungal recurrence. The patients were divided into three diagnostic categories: proven aspergillosis (autoptic or histologic diagnosis) n = 39, probable aspergillosis (radiological diagnosis with positive microbiology) n = 9, and possible aspergillosis (radiological diagnosis alone) n = 13. In the same period among 675 acute leukemia patients the incidence of proven or probable aspergillosis was 7.1%. At onset of infection 92% of patients were neutropenic (< 0.5 x 10(9)/L). The most frequent site of infection was the lung (90%); disseminated disease was present in 20 patients. Among 44 assessable patients, 12 (27%) failed to respond to early antifungal therapy and died. Thirty-two patients were cured with antifungal treatment, three of five nonneutropenic with only itraconazole, the others with amphotericin B 1 mg/Kg/day with or without itraconazole subsequently or with liposomal amphotericin, Ambisome, if renal toxicity occurred. Twenty-four of 29 neutropenic responders, all affected by acute leukemia, continued scheduled intensive chemotherapies. Pulmonary lobectomy was successfully combined with medical treatment in two cases before scheduled BMT. After infection nine patients were submitted to BMT (six allo, one marrow unrelated donor (MUD), two auto) with Ambisome or itraconazole as secondary prophylaxis without fungal relapse (follow-up: 25-99 months). The median time from fungal infection to transplant was five months, range 3-10. Thirteen of 29 surviving patients had leukemia relapse, but only three (23%) of these showed also fungal infection recurrence. In conclusion, a high index of suspicion and careful clinical and radiological examinations are the key to identifying infected patients early and to programming the following therapeutic steps. Above all in leukemia patients, prompt and aggressive administration of antifungal agents seems to improve the outcome of invasive fungal disease and to permit intensive chemotherapy completion and transplant.

Early addition of ribavirin to interferon in chronic hepatitis C not responsive to interferon monotherapy.

BACKGROUND/AIM: Persistence of HCV-RNA in serum early in treatment is a strong predictor of failure of alpha-interferon therapy for chronic hepatitis C. Therefore, we compared the efficacy of ribavirin addition to alpha-interferon with a doubling of the dosage of alpha-interferon in case of lack of early virological response to alpha-interferon therapy. METHODS: Sixty patients were administered interferon alpha2b at the dosage of 3 million units 3 times a week. After the first 4 weeks of therapy, serum HCV-RNA was evaluated. The patients with negative HCV-RNA test received the same treatment for a further 11 months, while those with detectable HCV-RNA were randomized to receive either the same dosage of alpha-interferon plus ribavirin (1000 mg/day) or double dosage of alpha-interferon (6 million units tiw) for 11 months. We considered sustained response to be the maintenance of normal alanine aminotransferase and negativity at HCV-RNA testing until the end of a 6-month post-treatment follow-up. RESULTS: After the first 4 weeks of treatment, 12 (20%) patients showed virological response and 48 patients (80%) remained positive on HCV-RNA testing. Sustained response was observed in 5/12 (42%) patients with early virological response, in 10/24 (42%) patients without early virological response who were administered ribavirin and alpha-interferon, and in only 1/24 (4%) patients who were administered the double dosage of alpha-interferon (p=0.006). CONCLUSIONS: This study shows the efficacy of the addition of ribavirin to alpha-interferon and the lack of efficacy of doubling the dosage of alpha-interferon in patients without clearance of hepatitis C virus early on in treatment.

Hepatitis C virus in non-Hodgkin's lymphoma. A reappraisal after a prospective case-control study of 300 patients. Lombart Study Group of HCV-Lymphoma.

It is widely thought, but not yet explained, that there might be a pathogenetic link between the infection of hepatitis C virus (HCV) and the onset of B non-Hodgkin's lymphoma (NHL). We studied the prevalence of serum anti-HCV antibodies among 300 NHL comparing it with the prevalence among 600 age- and sex-matched non-neoplastic subjects as controls, 247 patients with non-lymphomatous neoplasm, and 122 patients treated with immunosuppressive agents. We found a prevalence of 0.16 among NHL and 0.085 among controls and non-lymphomatous patients. Although the difference was statistically significant (P < 0.001), the odds ratio was 2.049 and its confidence intervals included the equality. The HCV prevalence was independent of NHL subset, and the genotypes distribution was the same among NHL and controls. We disclosed a HBsAg prevalence of 0.077 in NHL versus 0.008 in controls (P < 0.001) with an odds ratio of 9.9. We do not believe that these findings support the hypothesis of an HCV pathogenetic role in lymphomagenesis because (i) the risk of previous infection is marginally higher in NHL than in controls, (ii) a typical genotype distribution is lacking, as is a NHL clinico-histological feature associated with HCV, and (iii) the higher prevalence of viral infection is not specific as witnessed by the high HBsAg prevalence.

[The laboratory in the diagnosis of sexually transmitted diseases]. / Il laboratorio nella diagnosi delle malattie sessualmente trasmesse.

In light of the wide array of sexually transmitted infections, it is important that accurate, increasingly advanced laboratory procedures be used to identify the specific agents and that the results of these procedures be correctly interpreted. For this reason, guidelines for the laboratory diagnosis of sexually transmitted diseases (STD) must include standardised criteria and protocols. This report presents the results of several Italian studies on the variability of laboratory procedures for diagnosing the most common STD.

Should patients with early loss of serum HCV-RNA during alpha interferon therapy for chronic hepatitis C be treated for 6 or 12 months?

BACKGROUND/AIMS: Retrospective studies have suggested that early loss of serum HCV-RNA predicts sustained response to alpha-interferon treatment in chronic hepatitis C, but the optimal duration of therapy after loss of HCV-RNA is not known. The aims of this study were: a) to prospectively evaluate the effectiveness of HCV-RNA testing after 1 month of alpha-interferon treatment in the prediction of sustained response, and b) to compare the efficacy of 6 and 12 months of therapy in patients with a negative serum HCV-RNA test after the first month of treatment. METHODS: One hundred and thirty patients were administered interferon alpha-2b at doses related to body weight (< or > or = 60 kg) and to HCV genotype: 5 or 8 MU tiw for type 1, and 3 or 5 MU tiw for genotypes non-1. Serum HCV-RNA testing was performed using in-house nested RT-PCR at month 1, at the end of treatment and 6 months afterwards. We considered sustained response to be the maintenance of normal alanine aminotransferase and negativity at serum HCV-RNA testing until the end of follow-up. RESULTS: Sustained response was observed in 2/72 (2.8%) patients with detectable HCV-RNA after the initial month of therapy, in 8/30 (26.7%) patients with early loss of HCV-RNA treated for 6 months and in 20/28 (71.4%) patients treated for 12 months (p<0.01). CONCLUSIONS: Serum HCV-RNA detectability after the first month is strongly associated with a very poor chance of sustained response, and these cases should be offered other treatments. Patients with early loss of HCV-RNA should complete a 12-month treatment, which appeared more effective than a 6-month treatment.

Increased burn patient survival with once-a-day high dose teicoplanin and netilmicin. An Italian multicenter study.

This is the final report of a large, controlled, multicenter Italian study on immuno- and chemotherapy in adult patients with burns affecting 20 to 95% of total body surface area (mean 35%). The antibiotic treatment of burn patients consisted of topical silver sulfadiazine, short-term antimicrobial chemoprophylaxis with pefloxacin (800 mg i.v. qd) for the first 4 days and polychemotherapy with teicoplanin (800 mg i.v. qd) together with netilmicin (300 mg i.m. qd) in one or more cycles of 5-12 days. At random, half of the patients received thymostimulin, 70 mg i.m. qd for the first month and every other day thereafter. The analysis at completion of 634 valid cases showed that when the results are stratified by means of the Roi risk index, 396 of the 530 patients who contracted wound infection (84%) after chemoprophylaxis were in the first three categories and a mean of 95% survived. Of the remaining 134 patients (Roi index 4-5) only 50% survived. There was no difference in survival of the immunotherapy group in comparison with the parallel group without thymostimulin. The short-term antimicrobial prophylaxis prevented wound infection in only 104 of 634 patients (16%) and they were at low risk (84% Roi index 1). Of the bacterial pathogens involved in septic complications Staphylococcus aureus and Pseudomonas aeruginosa were prevalent (86%): eradication was achieved in 43% of patients and clinical cure or improvement were seen with combination chemotherapy in 64% of all patients, mainly with only one treatment cycle. This value increased to 79% for the 395 protocol-complying patients and went down to 20% in the 135 non-compliers. The total survival of complier and non-complier patients was 447 of the 530 valid patients (84%). The overall mortality of the 634 evaluable patients was 13.1%, ranging from less than 2% to 68%. Burn mortality was directly proportional to the percentage of burned body surface area, to increasing age and other variables of the Roi index, a 50% mortality being associated with a 72.5% total body surface area burned. Normoergic burn patients had a mortality rate of 9.1% versus 35.7% in anergic patients.

Efficacy of a three day course of azithromycin in moderately severe community-acquired pneumonia.

This study was designed to evaluate the efficacy of a 3 day course of azithromycin in low to moderately severe community-acquired pneumonia. Forty patients with low to moderately severe community-acquired pneumonia (29 males, 11 females, mean age 46 +/- 17 yrs; 20 pretreated with betalactams for 2-10 days with no results before admission to hospital; 18 with evidence of co-morbidity) were enrolled in an open, randomized study with azithromycin, 500 mg q.d. oral therapy for 3 days, versus clarithromycin, 250 mg b.i.d. oral therapy for 10 +/- 2 days. The aetiology of pneumonia was identified in 18 patients by serology (nine Mycoplasma pneumoniae, four Chlamydia pneumoniae, five Legionella pneumophila; one patient with chlamydial infection also had Klebsiella pneumoniae bacteraemia). A presumptive aetiological diagnosis was obtained with sputum culture in three other patients (one Haemophilus influenzae, two Haemophilus parainfluenzae), all strains were sole isolates with 10(8) Colony forming units (CFU), and with Gram stain in one patient with Streptococcus pneumoniae. All patients in the azithromycin group (one after a second 3 day course), and all but two (of those available for evaluation) of the clarithromycin group were cured. Defervescence occurred after 2.6 +/- 1.6 days, and chest roentgenogram cleared after 8.9 +/- 3.3 days, with no difference between the two groups. Tolerance was good, and there were no withdrawals from therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

Epidemiology of urogenital infections caused by Chlamydia trachomatis and outline of characteristic features of patients at risk.

A study of Chlamydia trachomatis infection was conducted in two stages on 15,656 subjects at urogenital clinics of the Faculty of Medicine and Surgery at La Sapienza University in Rome, the S. Anna Hospital in Turin, and the Niguarda Hospital in Milan. The overall incidence of the disease was 6.4% in patients examined throughout the whole study period. The rate of positive cases was 5.8% for the 5270 patients examined up to 1990, and 6.7% for the 10,386 patients examined from 1990 to 1992, showing an increasing trend. There was a much higher positivity rate in men (9.8%) than in women (6.0%); the difference was statistically significant. Of all patients, 60%, were asymptomatic. In symptomatic patients, C. trachomatis was present in 18.5% of cases of non-gonococcal urethritis and in 12.8% of cases of salpingitis. The highest incidence of C. trachomatis infection was in women who had begun sexual activity at an early age, (under 25 years in age), had several sexual partners and used intra-uterine contraceptive devices or spermicides or both.

Chlamydia pneumoniae infection in acute exacerbations of COPD.

Chlamydia pneumoniae, strain TWAR, is a frequent causative agent of acute respiratory disease. We assessed the incidence and prevalence of Chlamydia pneumoniae infections in COPD. We studied, from January 1990 to May 1991, 142 out-patients with acute purulent exacerbations of chronic obstructive pulmonary disease (COPD) and 114 healthy control subjects. Oropharyngeal swab specimens were collected at each exacerbation and analysed using a high definition monoclonal indirect fluorescent antibody test for Chlamydia pneumoniae identification. Immunoglobulins G and M (IgG and IgM) fractions of antibodies to Chlamydia pneumoniae were studied by microimmunofluorescence test. Prevalence of specific IgG was 63% in COPD, and 46% in controls (Chi-squared test p = 0.007). Moreover, mean titre of IgG was significantly higher in COPD than in controls. Five patients were positive for specific IgM (> or = = 1:16), and one had a fourfold increase of IgG titre; four of these patients had been treated with ciprofloxacin 1 g.day-1 for 10 days, and two with erythromycin, 3 g.day-1 for 14 days, with remission of signs and symptoms of exacerbation. Chlamydia pneumoniae identification was always negative. Our data suggest that Chlamydia pneumoniae infection is a rather frequent event in COPD, since at least 4% of exacerbations may be associated with it.

Updated evaluation of the activity of antibiotics in a burn centre.

The results of microbiological tests performed in a burns unit between January 1989 and December 1990 have been analysed. Burn wound swabs and biopsies, blood cultures, central venous and urinary catheters, bronchial aspirates, pharyngeal swabs and faecal cultures for a total of 7950 examinations were considered. Staph. aureus was the most frequently isolated bacterium, followed by Staph. epidermidis, Pseud. aeruginosa and E. coli. The antibiograms have shown a low efficacy rate of cephalosporins, even of the latest generation, while with Gram-positive isolates the highest rates of activity were recorded by vancomycin and teicoplanin (100 per cent sensitivity). The situation seems better with Gram-negative organisms since they appear to be sensitive to a larger number of antimicrobial agents.

A study of the incidence of urogenital Chlamydia trachomatis in patients attending specialized departments of Rome, Milan and Turin, Italy.

A study of Chlamydia trachomatis was conducted in 5270 subjects seen at clinics of the Faculty of Medicine and Surgery at La Sapienza University in Rome, the S. Anna Hospital in Turin, and the Provincial Maternity Hospital Institute in Milan. In these areas, C. trachomatis was present in 5.8% of the cases examined; in addition it was present with statistically significant frequencies in cases of salpingitis (49.1) and epididymitis (21.7). It may also be found in cases of extrauterine pregnancy, sterility and abortion. Those most affected were women who had begun their sexual activity at an early age, were under 25, had several sexual partners and who used the coil and/or spermicides. A routine check for C. trachomatis should be considered for those women with those risk factors.

Humoral circulatory immune response to Gardnerella vaginalis.

Strain-specific circulating immunoglobulin G and/or M was detected by enzyme-linked immunosorbent assay and immunofluorescence test by using Formol-treated suspensions of Gardnerella vaginalis from 28 women with overt vaginitis but only three symptom-free subjects among 43 otherwise healthy women found to be colonized by G. vaginalis. Analogous but less stringent strain specificity patterns were elicited by immunization of BALB/c mice.

[Chlamydia and pharyngotonsillitis in childhood]. / Chlamydia e faringotonsillite nell'infanzia.

The aim of this study was to evaluate the role of Chlamydia trachomatis in pharyngeal infections in childhood. 200 children, aged 1 month to 12 years, with and without symptoms of pharyngitis were examined for CT. Swabs were taken from the oropharynx and tonsillar areas for direct fluorescent antibody assay. 12% of the patients and 4% of the control subjects were positive for CT. Group A Streptococcus was not isolated from any patient or control subject positive for CT. In positive cases we extended the research to relatives. The treatment for positive cases was erythromycin and it eradicated CT in 80% of our patients. A new strain of Chlamydia, called TWAR, showing cross-reactivity with CT, has been isolated. Therefore further research should be carried out to discover if micro-organisms considered CT up to now, do belong or not to TWAR strain. Long term studies in children will be helpful in further defining the role of Chlamydia, trachomatis or TWAR, as a cause of pharyngitis.