Detection of genetic lineages of human metapneumovirus in Croatia during the winter season 2005/2006

Human metapneumovirus (HMPV) is an important respiratory pathogen, especially among young children. The genetic characteristics of HMPV circulating in Croatia have not been studied so far. The aim of this study was to determine the incidence of HMPV infection in hospitalized children with acute respiratory tract infection (ARTI) in the season 2005/2006 in Croatia, as well as to perform the genotypic analysis of detected HMPV strains. From December 1 to March 31 nasopharyngeal secretions (NPSs) were collected from 402 inpatients up to 5 years of age with ARTI. NPSs were tested by real-time RT-PCR assay targeting the nucleoprotein (N) gene of HMPV. HMPV infection was detected in 33 patients (8.2%). To perform the phylogenetic study, partial nucleotide sequences were obtained for HMPV fusion (F) gene of 30 HMPV positive samples. Phylogenetic analysis showed the circulation of two main genetic lineages (A and B), with B lineages being prevalent. It also showed the existence of two sublineages within the group B (B1 and B2) and three subclusters within lineage A (A1, A2a and A2b). Further molecular analysis revealed point mutations in HMPV strains of sublineage B1.     

The diagnostic value of atopy patch testing and prick testing in atopic dermatitis: facts and controversies

We conducted a systematic Medline search of the literature (1998-2008) on the criteria for performing the skin prick test and atopy patch testing (APT) to determine their utility in atopic dermatitis (AD). The skin prick, scratch, and skin patch tests are performed to identify which allergen is causing eczematous skin symptoms in patients with AD, or sneezing, nasal congestion, itchy eyes, wheezing, skin rash, and swelling. Many allergens in foods, drugs, and environmental substances (eg, ragweed and fungus), as well as contact allergens, can elicit eczematous skin reactions after epicutaneous application. Because no gold standard exists for aeroallergen provocation in AD, the APT is currently used to evaluate allergen without comparison with another accurate and reliable method. The APT is presumed to reflect delayed-phase clinical reactions. Even with delayed onset of symptoms (more than 2 hours after food ingestion), APT findings were not consistent among AD children. The APT could be used in children with gastrointestinal reactions to foods as well as AD. After standardization, the APT may provide further diagnostic information in addition to the skin prick test and serum immunoglobulin E values and may be able to evaluate the actual clinical relevance of immunoglobulin E-mediated sensitizations for eczematous lesions. The European APT model used with standardization of allergen concentration and vehicle may provide an important diagnostic tool to select patients for avoidance and for procedures of allergen-specific immunotherapy, but the clinical relevance of positive APT reactions awaits standardized provocation and avoidance testing.     

Online on-the-spot searching increases use of evidence during consultations in family practice

OBJECTIVE: The project aimed to search for online evidence in a structured way in consultation with the patient, to investigate whether the evidence discovered changed decisions. METHODS: We developed the "Online on-the-spot" method (OOS) as a part of a quality improvement program. Within a general practice consultation three physicians and two trainees searched in a fixed pattern and sequence the national guidelines of general practitioners developed by the Dutch College of General Practitioners, Clinical Evidence, Trip-database and the British Medical Journal. All GPs who performed this quality improvement program were in favor of the project. RESULTS: During 3 months five GPs registered 365 searches out of 2920 patient-doctor contacts. For each eight patient-doctor contacts there was one online search. Patients were actively involved in 53% of the searches (95%C.I.: 48-57%). On average, two databases were consulted. An answer to the question was found in 87% of cases and in almost half of cases it was relevant new information for the doctor. The GP changed his decision due to the problem in 26% (95%C.I.: 21-29) of cases. At the end of the OOS project, the number of searches within 5 min were significantly higher than at the start: 51% (95% C.I.: 44-59) to 33% (95% C.I.: 24-43), respectively. CONCLUSIONS: The OOS project is a timely answer to the doctors' educational needs in attending to the patient. PRACTICE IMPLICATIONS: OOS could connect the patient, the doctor and the evidence.     

Exploring Equisetum arvense L., Equisetum ramosissimum L. and Equisetum telmateia L. as sources of natural antioxidants

The antioxidant and scavenging activities of above ground parts of Equisetum arvense L., Equisetum ramosissimum L. and Equisetum telmateia L. phosphate buffer (pH 7) extracts were investigated. Activities of antioxidant enzymes (superoxide dismutase, catalase, guaiacol peroxidase and glutathione peroxidase), quantities of reduced glutathione, malonyldialdehyde, superoxide and hydroxyl radicals and flavonoid, soluble protein, chlorophyll a, b and carotenoid contents were determined. The total antioxidant capacity was determined by ferric reducing antioxidant power (FRAP) assay. The Equisetum telmateia extract demonstrated scavenging and antioxidant properties better than Equisetum ramosissimum and Equisetum arvense. The ESR signal of DMPO‐OH radical adducts in the presence of Equisetum telmateia phosphate buffer (pH 7) extract was reduced by 98.9% indicating that Equisetum telmateia could be a useful source of antioxidants with huge scavenging ability. Copyright © 2008 John Wiley & Sons, Ltd.     

Carboplatin resistant human laryngeal carcinoma cells are cross resistant to curcumin due to reduced curcumin accumulation

Curcumin is a natural compound that exhibits a wide range of beneficial effects, among them the anti-tumor activity. Recently it was shown that curcumin may be efficient against drug resistant tumor cells. The goal of our investigation was to examine if human laryngeal carcinoma cells resistant to carboplatin display sensitivity to curcumin, as compared to parental cells, and if this sensitivity is altered, to determine the molecular mechanisms that are responsible for it. We found that carboplatin resistant 7T cells were also cross resistant to curcumin. After the treatment with equimolar concentration of curcumin, 7T cells exhibited lower intracellular accumulation of curcumin which coincided with reduced formation of reactive oxygen species (ROS), diminished lipid and DNA damage followed by reduced induction of apoptosis and expression of heat shock protein 70 (Hsp70), as compared to parental HEp-2 cells. However, after the treatment with equitoxic concentration of curcumin, intracellular accumulation and all the explored downstream effects were similar in both cell lines suggesting that resistance of 7T cells to curcumin was based on its reduced intracellular accumulation. Since curcumin accumulates mostly in the membranes, we explored the fatty acid composition of both cell lines, but we did not find any difference between them.     

Intrauterine inflammation and the onset of peri-intraventricular hemorrhage in premature infants

BACKGROUND: Peri-intraventricular hemorrhage (P/IVH) is a common neonatal morbidity among premature infants. The aim of the study was to examine the association between placental and/or fetal inflammation and the onset of P/IVH in premature infants. METHODS: A prospective study included 125 infants with gestational age 23-29 weeks. Placentas were examined for the presence of chorioamnionitis and funisitis, cord blood was sampled for the measurement of cytokines (IL-6 and IL-8). Fetal inflammation was defined as levels of IL-6 higher than 7.6 pg/ml. P/IVH was defined as early if diagnosed within the 1st day after birth; thereafter P/IVH was defined as late. RESULTS: Adjusted for the influence of gestational age, early-onset sepsis (OR 3.2, p = 0.045) and no or incomplete antenatal steroid course (OR 6.0, p = 0.001) significantly predicted early P/IVH. Funisitis (OR 1.6, p = 0.06) and fetal inflammation (OR 2.6, p = 0.06) were only partially associated with early hemorrhage. Contrary to that, respiratory distress syndrome (OR 3.4, p = 0.04), mechanical ventilation (OR 5.9, p = 0.008), low blood pressure (OR 3.5, p = 0.02), and vasopressors (OR 5.7, p = 0.002) were associated with late P/IVH. In multivariate analysis no or incomplete steroid course remained independent predictors for early and use of vasopressors for late P/IVH. The interaction of fetal inflammation and vaginal delivery with no or incomplete steroid course increased the risk of early P/IVH. CONCLUSIONS: These results indicate different risk factors for early and late P/IVH. Neither funisitis nor fetal inflammation independently predicts the onset of P/IVH. However, the interaction of fetal inflammation and vaginal delivery with no or incomplete antenatal steroid course increase the risk of early but not also late P/IVH.