Isolation and genotyping of Acanthamoeba strains from corneal infections in Italy.

Acanthamoeba keratitis (AK) is a corneal disease caused by members of a genus of free-living amoebae and is associated predominantly with contact lens (CL) use. This study reports 16 cases of culture-proven AK diagnosed in northern Italy. Genotype identification was carried out with a PCR assay based on sequence analysis of the 18S rRNA gene, and sensitivity and specificity were evaluated in comparison with traditional parasitological techniques. A 405 bp region of the 18S rRNA gene (ASA.S1) including diagnostic fragment 3 (DF3) was amplified using the genus-specific primers JDP1 and JDP2. Genotype assignment was based on phenetic analysis of the ASA.S1 subset of the nuclear small-subunit rRNA gene sequence excluding the highly variable DF3 region. Phylogenetic analysis was also performed on the sequences obtained. All patients complained of monolateral infection; 11 (68.75%) admitted improper CL disinfection. In 14/16 (87.5 %) subjects, corneal scrapings were stained with calcofluor white and haematoxylin and eosin and, in ten cases (62.5 %), microscopy was positive for Acanthamoeba cysts. In vitro culture on 3 % non-nutrient agar plates was obtained in all cases (100 %), whereas cloning and axenic growth were positive for 14 amoebic stocks (87.5 %). PCR analysis had 100 % sensitivity and specificity compared with in vitro axenic culture, showing positive amplification from 15 isolates. All Acanthamoeba strains belonged to the T4 genotype, the main AK-related genotype worldwide. These results confirmed the importance of a complete diagnostic protocol, including a PCR assay, for the clinical diagnosis of AK on biological samples. Genotyping allowed inclusion of all isolates in the T4 group, thus demonstrating the prevalence of this genotype in northern Italy.

Predictive role of single diseases and their combination on recovery of balance and gait in disabled elderly patients.

OBJECTIVES: In the elderly population, chronic diseases are common determinants of mobility limitations and comorbidity consistently shows a strong association with functional status. This study was designed to evaluate the role of single chronic diseases and of their combination on functional recovery after rehabilitative treatment in disabled elderly patients. DESIGN: With respect to the difference in magnitude of their disabling effect, diseases were classified into 2 groups: "more disabling" diseases (COPD, heart failure, peripheral artery diseases, diabetes, and not life-threatening cancer) and "less disabling" diseases (anemia, kidney, gastrointestinal, and liver diseases). SETTING: 35-bed Geriatric Evaluation and Rehabilitation Unit. PARTICIPANTS: We studied 710 patients (age 77.8 +/- 7.4 years, 76.2% females), consecutively admitted for stroke, Parkinson's disease, and osteoarthritis. MEASUREMENTS: A multidimensional evaluation for mobility (Tinetti-score), cognitive status (MMSE), and somatic health (Greenfield's Individual Disease Severity Index-IDS, Burden of diseases-BoD) was performed. Functional recovery was decided based on the Delta-Tinetti, which is the difference of the values between admission and discharge. RESULTS: We tested, in a multivariate regression model, the predictive role of single chronic conditions and of their combinations on functional recovery, after having adjusted for which diseases are direct causes of disability (stroke, Parkinson's disease, and osteoarthritis) and other potential predictors (age, sex, cognitive function, depressive symptoms, albumin, and c-reactive protein). A negative prediction of functional recovery was expressed by the "more disabling" diseases group. The determinants of poor recovery were characterized by the combination of "more disabling diseases" rather than single condition effects, independently by age, cognitive, and functional status on admission. CONCLUSION: Our study adds a new perspective about the role of COPD, heart failure, peripheral artery diseases, diabetes and not life-threatening cancer on functional recovery, emphasizing their combined impact in elderly people.

Diagnosis of chronic pain caused by osteoarthritis and prescription of analgesics in patients with cognitive impairment.

OBJECTIVE: Evaluation of the ability to detect symptoms of osteoarthritis and prescription of analgesics in older adults with different degrees of cognitive impairment. SETTING AND PATIENTS: Patients consecutively admitted to a Geriatric Evaluation and Rehabilitation Unit (GERU) for disability due to orthopedic, neurological, or cardiopulmonary diseases, after a surgical intervention, or for behavioral disturbances of dementia. Subjects with a recent orthopedic intervention were excluded. Eight hundred eighty-eight patients were analyzed (mean age 78.4 +/- 7.2 years, 69.5% female) and divided into 4 groups according to cognitive impairment: severe (Mini-Mental State Examination, MMSE 0-12), moderate (MMSE 13-18), mild (MMSE 19-24), or absent (MMSE 25-30). METHODS: Patients underwent a geriatric multidimensional assessment; the diagnosis of musculoskeletal pain was based on patients' direct report, objective signs (inflammatory signs around the joints), and "pain behaviors" (reductions in activity, social withdrawal, self-protective maneuvers, increased alterations in facial expressions or body postures, observable displays of distress). Analgesic drugs were registered on discharge in subjects with symptomatic osteoarthritis. RESULTS: Of the patients admitted to GERU, 16.8% (n = 149) had osteoarthritis; this percentage is different according to cognitive impairment (MMSE 0-12: 1%, MMSE 13-18: 8%, MMSE 19-24: 17.4%, MMSE 24-30: 34.1%, P < .001). If patients admitted for osteoarthritis are excluded, the new diagnoses of pain due to osteoarthritis are 306 (41.4%), without differences between cognitive status groups. On discharge, analgesic drugs are prescribed to 28.8% of patients with symptomatic arthritis (n = 131); the percentage is lower in patients with severe cognitive impairment in comparison with subjects with better cognitive functions. CONCLUSION: There is a poor attention to chronic pain due to osteoarthritis in community-dwelling elders with cognitive impairment. In a rehabilitative setting, an accurate evaluation of objective signs allows the diagnosis of symptomatic osteoarthritis. However, the link between the diagnosis and the treatment is not direct, since the prescription of analgesics is lower in patients with a more severe cognitive impairment.

Fatal Naegleria fowleri meningoencephalitis, Italy.

We report the first case of primary amebic meningoencephalitis in Italy, in a 9-year-old boy. Clinical course was fulminant, and diagnosis was made by identifying amebas in stained brain sections and by indirect immunofluorescence analysis. Naegleria fowleri was characterized as genotype I on the basis of polymerase chain reaction test results.

Amebic infections due to the Entamoeba histolytica-Entamoeba dispar complex: a study of the incidence in a remote rural area of Ecuador.

An epidemiologic field study was conducted in the village of Borbòn in Esmeraldas province in northern Ecuador to compare different parasitologic methods in the diagnosis of infection with the Entamoeba histolytica/Entamoeba dispar complex. The results of two stool antigen detection assays (the Prospect Entamoeba histolytica microplate assay and the E. histolytica II assay) were compared with isoenzyme characterization of the amebic isolates. Nearly all (176 of 178, 98.9%) subjects were positive for intestinal parasites on direct microscopic examination, and cysts and/or vegetative forms morphologically consistent with the E. histolytica/E. dispar complex were recorded in 48 of 178 cases (27%). Culture in Robinson's medium was positive for amebic stocks in 89 (50%) of the 178 samples tested. Of the 37 isolates successfully stabilized, cloned, and characterized by zymodeme analysis, seven (18.9%) showed isoenzyme patterns of E. histolytica, whereas 26 (70.3%) showed patterns of E. dispar. The remaining four strains were identified as Entamoeba coli (three isolates; 8.1%) and Dientamoeba fragilis (one strain; 2.7%).The immunochromatographic tests showed different degrees of sensitivity and specificity when compared with isoenzyme characterization as the reference technique. The microplate assay, which does not discriminate between E. histolytica and E.dispar, showed a sensitivity of 54.5% and a specificity of 94% for both these amebic species. In contrast, the second-generation E. histolytica II test had a sensitivity of 14.3% and a specificity of 98.4% for E. histolytica sensu stricto. Our survey clearly demonstrated that more specific and sensitive diagnostic tests, such as stool antigen detection assays and isoenzyme analysis, are needed to establish the actual worldwide distribution of E. histolytica and E. dispar.