Microbiological investigations on a nosocomial case of Legionella pneumophila pneumonia associated with water birth and review of neonatal cases.

A case of Legionella pneumophila 1 pneumonia, confirmed by positive serology and urinary antigen, occurred in a 7-day old neonate after water birth in hospital. As respiratory samples were not available for culture, further microbiological investigations were performed in neonate and environment, in order to recognize the source of infection. The hospital water supply was contaminated by L. pneumophila 1 strains (300-2000 cfu/L) of two monoclonal subtypes of Pontiac subgroup. L. spiritensis (10-225 cfu/L) was isolated from cold tap water of the patient's home. PCR from tap and humidifiers water of the patient's home was positive for Legionella spp, but not for L. pneumophila. Because L. pneumophila 1, responsible of child infection, was only isolated from the hospital pool water for waterbirthing, we conclude that the infant acquired the nosocomial legionellosis by prolonged delivery in contaminated water, perhaps by aspiration. Infection control measures for waterbirthing are highly recommended. A review of neonatal case of legionellosis is also presented. As this rare infection may have a high fatality rate if unrecognized, pediatricians should be aware of the possibility of the legionellosis in newborns.

Clarithromycin and amoxicillin susceptibility of Helicobacter pylori strains isolated from adult patients with gastric or duodenal ulcer in Italy.

Helicobacter pylori strains, isolated from 100 gastric biopsies from 49 previously untreated adult patients with endoscopy and histology-confirmed gastric or duodenal ulcer, were tested for in vitro antimicrobial susceptibility. Strains were isolated from biopsies of 75.5% (37 of 49) patients before therapy and of 13.5% after therapy. Clarithromycin and amoxicillin susceptibility testing was performed on pretreatment and posttreatment strains by using the agar disk diffusion method and E-test, a quantitative technique for the minimal inhibitory concentration (MIC) determination. All strains (n = 53) were susceptible to amoxicillin by the two methods. Three strains of 34 (8.8%) patients were resistant to clarithromycin: two by both methods and one by E-test (MIC > 2 microg/ml). E-test, although more expensive than the disk diffusion method, is easy to perform and is a reliable method for testing H. pylori susceptibility to antimicrobial agents in the clinical microbiology laboratory.

Is the clinical course of HIV-1 changing? Cohort study.

OBJECTIVE: To assess whether the clinical course of HIV infection has changed from 1985 to 1995. DESIGN: Cohort Study. SETTING: Infectious disease clinic. SUBJECTS: 285 patients recruited from September 1985 to January 1995 with < or = 12 months between the dates of their last seronegative and first seropositive test result and with first follow up visit in the six months after seroconversion and at least 12 months' follow up. Patients were grouped according to the date of seroconversion. MAIN OUTCOME MEASURES: Time to CD4 cell count of < 500, 400, and 200 x 10(6) cells/l, and clinical outcome defining AIDS; variation in cell count per day between consecutive visits, and ratio between this variation and time from estimated date of seroconversion at each visit. RESULTS: The groups were similar in age, number with acute primary HIV infection, CD4 cell count at intake, and cell count at the beginning of antiretroviral treatment; they differed in sex ratio, risk factors for HIV, probability of CD4 cell decline to < 500, 400, and 200 x 10(6) cells/l. and risk of developing AIDS. Acute infection, seroconversion after December 1989, and serum beta 2 microglobulin > 296 nmol/l were independent predictors of poor clinical course. The speed of CD4 cell decline, expressed as cell variation divided by the number of days between consecutive visits, increased with more recent seroconversion (P = 0.02). Ratio between the speed of CD4 cell decline and time from estimated date of seroconversion at each visit was also higher in the patients who seroconverted after December 1989. CONCLUSIONS: The faster disease progression and the higher speed of CD4 cell decline at early stages in the patients with recently acquired HIV infection suggest changes in the clinical course of HIV infection.

Phytosterol compounds having antiviral efficacy.

The present study is focused on the antiviral action patterns obtained in vitro with synthetic sterolester comprising compositions on virus-bearing host cell-lines. Appropriate cell-lines were infected with HIV-1, human Cytomegalovirus (HCMV) and Herpes simplex virus (HSV). There appears to exist a clear anti-infective efficacy for a selected number of such ester compounds, provided they are formulated into spontaneously dispersible concentrates, which in aqueous dilution engender ultramicro-emulsions having micelles in the lowest nanosize region. A significant protection against HIV-induced cytopathogenic effect was demonstrated employing a methyltetrazolium salt reduction assay on HIV-infected MT4 cells when they were incubated with such concentrates. A similar effect was evidenced with the same concentrates, when preincubating concentrated virus, but not the target cells. Antiviral activity appeared to be remarkable also on HCMV infections in vitro, where a blocking effect on immediate-early antigen expression in fibroblast monolayers could be observed. Similarly, HSV-associated glycoprotein antigen in VERO cells also suggests that virus-cell interaction and/or virus multiplication could have been blocked at a very early point of time. This would be quite different from antiviral action-patterns studied so far and imputed into the current models of explanation. Proper solubilization of the employed phytosterol compounds is essential for achieving the described activity modes. The often recommended liposome formulations would not be well suited for such compounds and such purpose, since after dilution they produce aqueous macro-emulsions, only. Furthermore, liposome formulations tend to coalesce and exhibit Marangoni effects.

Coinfection and superinfection of hepatitis B virus in patients infected with human immunodeficiency virus: no evidence of faster progression to AIDS.

The influence of hepatitis B virus (HBV) on the natural history of human immunodeficiency virus (HIV) infection was evaluated in a prospective study of 347 HIV-positive, AIDS-free individuals infected through injecting drug use and sex and with known seroconversion dates. End points were CD4+ cell count < 200 x 10(6) cell/L and AIDS diagnosis. At entry, 229 had seromarkers to HBV; during the study, 107 had a CD4+ cell count < 200 x 10(6) cells/L and 66 developed AIDS. HBsAg chronic carriers, HBV infection-free subjects and those with baseline evidence of prior HBV infection did not differ in rates of progression to end points. Sexual transmission of HIV was significant predictor of CD4+ cell decline to < 200 x 10(6) cells/l [Hazard ratio (HZ): 1.56, 95% confidence interval (CI): 1.06-2.29, p = 0.0232] and progression to AIDS (HZ: 1.91, CI: 1.17-3.11, p = 0.0091). 15 HIV-positive and HBV infection-free patients had HBV seroconversion. They did not differ from those who remained HBV infection-free in rates of progression to end points, but 40% of them became HBsAg chronic carriers. These results suggest that HBV has no influence on progression of HIV disease, but that patients who have HIV before their HBV infection are more likely to become HBsAg chronic carriers than those who are infected with HBV before HIV.

Vulvodynia and HIV: causal or casual association?

BACKGROUND: No study exists on HIV and vulvodynia. OBJECTIVE: To evaluate vulvodynia in HIV infected women and its possible association with HIV. DESIGN: Cross sectional study. SETTING: Department of Medical and Surgical Sciences, Section of Infectious Diseases, University of Turin. METHODS: 235 HIV positive and 272 HIV negative women were studied for vulvar pain between June 1995 and January 1997. Diagnosis of vulvodynia was based on clinical history, dermatological and vulvovaginal examination, and bacteriology. Colposcopy and vulvar biopsy were performed and psychometric examination was done. RESULTS: Vulvodynia was diagnosed in five women with HIV and in one uninfected woman (odds ratio = 2.5; 95% confidence interval: 0.1-62.6). High neuroticism scores were observed in women with history of vulvar pain, the highest in HIV infected vulvodynia patients (p = 0.000). CONCLUSIONS: Confirming the association of gynaecological pain with neurotic personality, our study suggests causal link between HIV and vulvodynia.

[Atypical hepatitis]. / L'epatite atipica.

Acute viral hepatitis has a rather homogeneous clinical pattern, excepting a few features which are related to the specific viral agent. However, a few cases may be defined "atypical", as fas as clinical features, transmission pattern, serology are concerned. These include cholestatic hepatitis syndrome, fulminant HEV hepatitis during pregnancy, neonatal hepatitis. Non-typical aspects may be observed when extrahepatic manifestations are present (cryoglobulinemia, bone marrow aplasia, guillain-Barré syndrome, peripheral neuropathy, skin involvement, etc.). Finally, viral hepatitis may be "atypical" when associated with other infections (malaria, typhoid fever, etc.), mostly in patients from tropical Countries.

Tracheobronchial aspergillosis in a patient with AIDS treated with aerosolized amphotericin B combined with itraconazole.

The clinical features of a tracheobronchial infection due to Aspergillus flavus in an AIDS patient with a normal neutrophil count is described. Diagnosis was made by culture and microscopic examination of biopsies obtained from bronchial vegetations seen at bronchoscopy. Radiographic examination of the neck revealed the presence of large endoluminal fungal masses. Initially the patient was treated with a combination of itraconazole, flucytosine and aerosolized amphotericin B, then only with itraconazole plus aerosolized amphotericin B. A good therapeutic response was observed.

Serum cytokine profiles in acute primary HIV-1 infection and in infectious mononucleosis.

Serum cytokine profiles, T-cell subsets, and general parameters of immune activation were evaluated in 15 patients with acute primary HIV-1 infection, and compared with those obtained from 18 patients with acute primary Epstein-Barr virus (EBV) infection and from 18 control subjects in order to elucidate possible defects of immune response to HIV in early phases of virus-host interaction. Mean CD4+ cell count, serum concentrations of interleukin (IL)-2, IL-4, soluble IL-2 receptor (sIL-2R), tumor necrosis factor (TNF)-alpha, 5'-neopterin, and beta 2-microglobulin were significantly lower in acute HIV-1 infection than in EBV infection. Both acute HIV-1 and EBV infections were characterized by significantly higher mean CD8+ cell count and soluble CD8 antigen (sCD8) levels compared to control subjects, while acute HIV-1 infection was accompanied by the highest interferon (IFN)-gamma serum concentrations. In primary HIV-1 infection, significant impairment of CD4+- mediated T-helper function may lead to viral escape and persistence of infection despite an early and vigorous CD8+ T-lymphocyte activation.

Altered adrenocorticotropin and cortisol response to corticotropin-releasing hormone in HIV-1 infection.

Alterations of the hypothalamic-pituitary-adrenal (HPA) axis are common in HIV infection. To characterize further the site of these derangements and their possible causes, eight male drug addicts with symptomatic HIV infection (stage IV C2) underwent the following investigations: repeated baseline determinations of cortisol, adrenocorticotropin (ACTH), interleukin 1 beta (IL-1 beta), IL-6 and interferon alpha (IFN-alpha); and ovine corticotropin-releasing hormone (CRH) test (100 micrograms IV) for ACTH and cortisol determinations. Baseline cortisol levels were either normal or elevated in all patients. A significant linear correlation was found between baseline levels of cortisol and both IL-6 (r = 0.955; p < 0.001) and IL-1 beta (r = 0.863; p < 0.005), but not between cortisol and ACTH or between ACTH and circulating cytokines. Both ACTH and cortisol responses to CRH were nearly absent in six out of eight patients, and delayed in the others. The areas under the curves of both ACTH and cortisol after CRH were significantly lower in HIV patients than in a group of eight healthy control subjects (p = 0.0157 for ACTH and p = 0.046 for cortisol). Out data suggest the possibility of an inappropriate stimulation of the HPA axis in symptomatic HIV infection by HIV-induced release of cytokines, with a blunted pituitary and adrenal response to CRH.

Seroprevalence, risk factors and attitude to HIV-1 in a representative sample of lesbians in Turin.

OBJECTIVE: To estimate the HIV-1 seroprevalence, behavioural risks and attitude to HIV-1 infection among lesbians. SETTING: Institute of Infectious Diseases, University of Turin, Italy. SUBJECTS: From March 1992 to May 1993, 181 lesbians were tested for HIV-1 and included in the study. METHODS: Sociodemographic details, nonsexual risks for HIV-1, sexual behaviour, STD history and attitude to HIV-1 were obtained from an anonymous, standardised, self-administered, 30-item questionnaire. Snow-ball techniques were used to recruit the largest possible number of participants. RESULTS: 11 lesbians (6.1%) were found to be HIV-1 antibody positive. Of them, 10 were intravenous (i.v.) drug users. STD episodes were higher among lesbians with HIV-1 than without (p = 0.04), increasing in both groups over time. Syphilis, genital herpes and viral hepatitis were highly associated with HIV-1 (p = 0.000). In univariate analysis, i.v. drug use, bisexual behaviour, history of STDs, sex during menses and vaginal/anal manipulation were significantly linked to HIV-1 (p = 0.000). In multivariate analysis only history of i.v. drug use (p = 0.04) and bisexual behaviour (p = 0.06) remained independently associated with HIV-1. Seventy-one participants (39.3%) had already undergone AIDS testing. Only 3.5% admitted to be at risk for HIV-1 and 11% changed their sex habits after first hearing of AIDS. No lesbian had ever practised safe-sex. Television was the most important source of information on HIV-1 (84%). CONCLUSION: I.v. drug use was the most likely means of HIV-1 infecting the lesbians of Turin. The high rate of STDs and the low perceived risk to HIV-1 require programmes of STD prevention and AIDS information to be targeted at the lesbian community.

Rapid diagnosis of malaria by use of fluorescent probes.

Malaria diagnosis relies on observation of parasites in blood smears and the Giemsa-stained thick blood smear (G-TS) is the reference test. Diagnosis by G-TS in low-density infections requires long periods of observation and experienced microscopists. Examination of Giemsa-stained thin smears enables more reliable differentiation of species but may miss low-grade infections. Fluorescent stains may offer an alternative technique. We compared the Giemsa technique with 4,6-diamidine-2-phenilindolo-propidium iodide (DAPI-PI) stainings in order to evaluate the time required for diagnosis. A Plasmodium falciparum-infected blood specimen was diluted to obtain concentrations ranging from 6192 to 24 parasites/microliters (p/microliter), and thin and thick smears were stained with the two methods. The DAPI-PI proved useful: parasites were easily recognized and their morphology was preserved in thin and thick smears. The method allowed more rapid evaluation of thin smears as compared with Giemsa staining and enabled recognition of parasites in case of low-level parasitemias. The DAPI-PI staining technique may acquire an important role in malaria diagnosis, especially in nonendemic countries where technicians are not experienced with G-TS; in developing countries, it could be used in epidemiologic surveys of populations with low-density parasitemias, for which it enables a fast examination of smears and possibly the identification of parasite species.

Cytokine network and acute primary HIV-1 infection.

OBJECTIVE: To investigate the relationship between cytokine serum levels, peripheral blood lymphocyte subsets and clinical picture in acute primary HIV-1 infection. PATIENTS AND METHODS: Absolute number/microliters total lymphocytes, CD4+, CD8+ and natural killer (NK) cells, as well as serum levels of soluble CD8 receptor, interleukin (IL)-1 beta, IL-2, IL-4, IL-6, tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, beta 2-microglobulin and 5'-neopterin were determined in 15 patients with acute primary HIV-1 infection, 16 asymptomatic HIV-1-seropositive individuals and 18 HIV-1-seronegative individuals at risk for HIV-1 infection. RESULTS: Acute primary HIV-1 infection was characterized by significant CD4+ lymphocytopenia with low IL-2 serum concentrations, and by high absolute number of circulating CD8+ and NK cells, with elevated serum levels of soluble CD8 receptor, IL-1 beta, IFN-gamma and 5'-neopterin. Follow-up of acute seroconverters showed a significant decrease in NK cell counts and IL-1 beta levels, with an increase of IL-6. CONCLUSIONS: In acute primary HIV-1 infection, significant alteration of cytokine release, possibly induced by viral antigens, could be responsible for both clinical picture and activation of cytotoxic cells through abnormal mechanisms.

Risk of developing AIDS after primary acute HIV-1 infection.

We enrolled 134 newly human immunodeficiency virus type 1 (HIV-1)-infected subjects in a prospective study to determine the natural history of the infection and evaluate the risk of developing AIDS after acute primary HIV-1 infection (API). Twenty-three patients were observed during an acute primary HIV-1 infection, and 111 were asymptomatic seroconverters. Acute primary HIV-1 infection was more frequently observed in subjects who had acquired the infection through sexual transmission. Intravenous drug users were rarely affected and presented with milder symptomatology. Patients observed with an acute primary HIV-1 infection had a significantly higher risk of developing AIDS than asymptomatic seroconverters (68% at 56 months vs. 20% at 66 months; p = 0.026). Low CD4+ cell counts at the onset of acute illness and delayed seroconversion in enzyme-linked immunosorbent assay (ELISA) were associated with evolution to AIDS in acute seroconverters (p = 0.03 and 0.02, respectively). During the follow-up, patients with an acute illness were more likely to show an early fall of CD4+ cell counts below 200/microliters than asymptomatic seroconverters. The results of this follow-up study suggest the opportunity to study antiviral treatment protocols in patients with API as a possible measure to control disease progression.

Meningoencephalitis caused by Toxoplasma gondii diagnosed by isolation from cerebrospinal fluid in an HIV-positive patient.

Toxoplasmosis of the central nervous system is a frequent opportunistic infection in AIDS patients, usually presenting as a mass brain lesion detected by computerized axial tomography scanning or magnetic resonance imaging. A case of diffuse meningoencephalitis with no radiological evidence of brain lesions is described. Diagnosis was made by culturing cerebrospinal fluid (CSF) on THP1 cells where tachyzoites of Toxoplasma gondii were demonstrated after 8 days of incubation by both direct observation and immunofluorescence. CSF examination with culture should be considered in AIDS patients with neurological signs and symptoms but without radiological evidence of cerebral lesions.

Growth of Yersinia species in artificially contaminated blood bags.

Transfusion blood bags were inoculated with less than 0.1 colony-forming units (CFU) of Yersinia enterocolitica (serotypes O:3, O:5, and O:9) and Yersinia pseudotuberculosis I per mL and stored at 4 degrees C for 41 days. During storage, samples were collected periodically and colony counts of serial dilutions were performed by plating on tryptic soy agar. After a lag-phase, the growth of Yersinia species from less than 0.1 CFU per mL to 10(9) CFU per mL was observed in 3 of 10 bags. The data showed that prolonged storage of contaminated blood bags allows Y. enterocolitica and Y. pseudotuberculosis to multiply to high levels.