Stabilization of asthma prevalence among adolescents and increase among schoolchildren (ISAAC phases I and III) in Spain

BACKGROUND: Most studies show a steep increase in asthma prevalence in the last decades, although few studies had applied the same methodology. Recent reports point out the possibility that the epidemic has come to an end. We have studied the prevalence of asthma in a very large sample of children, repeating the study eight years apart. METHODS: Repeated cross-sectional studies using the International Study of Asthma and Allergies in Childhood (ISAAC) protocol in a sample of Spanish schoolchildren 6-7 (parent-reported) and 13-14 (self-reported) years old in 1994-95 (phase I) and 2002-2003 (phase III). The number of participants was 42 417 in phase I and 42 813 in phase III. The participation rate was over 87% (13-14 years) and 70% (6-7 years). RESULTS: The prevalence of wheezing in the previous year in children aged 13-14 years was 9.0 and 9.3% for boys and 9.6 and 9.2% for girls for phases I and III, respectively. Children 6-7 years of age showed a substantial increase in wheezing in the previous year (7.0 and 10.7% for boys and 5.3 and 8.2% for girls). Other symptoms and severity indexes followed the same patterns. CONCLUSIONS: In the last 8 years, the prevalence of asthma has not changed in 13-14-year-old Spanish children but has increased substantially in 6-7-year olds.     

Regional lymphadenopathy in cat-scratch disease: ultrasonographic findings

Background. Cat-scratch disease (CSD) is considered to be an emerging disease worldwide and is caused by Bartonella henselae, a gram-negative bacterium introduced by a scratch or bite of a cat. The most common clinical manifestation is regional lymphadenopathy, but clinical recognition may be difficult, as atypical manifestations may occur. The diagnosis is confirmed with serologic testing and histology is rarely needed. This paper is based on our experience with the use of ultrasonography in the diagnosis of CSD.¶Objective. The aim of this study was to describe the sonographic and color Doppler appearances of regional lymphadenopathy in CSD, as this has not widely reported in the literature. ¶Materials and methods. Forty-seven patients (average 9.4 years) were included who all had serologically and/or histologically proven CSD and had been studied using US early in the clinical course. All had a positive history of exposure to cats and exhibited regional lymphadenopathy.¶Results. US showed large hypoechoic adenopathy with some transmission enhancement and high vascularization on color-flow Doppler imaging. In 30 patients, abdominal US was also performed and splenic and/or hepatic granulomata were found in 10.¶Conclusions. In our experience, sonography and especially color-Doppler and power-Doppler sonography was helpful in the diagnosis of CSD. We believe it should be used in the initial study of children with regional lymphadenopathy, and serologic testing should be performed when CSD is suspected.     

Enteropathogenic Escherichia coli strains carrying genes encoding the PER-2 and TEM-116 extended-spectrum beta-lactamases isolated from children with diarrhea in Uruguay

We studied 13 extended-spectrum beta-lactamase (ESBL)-producing enteropathogenic Escherichia coli isolates from children suffering acute diarrhea in Uruguay. ESBL characterization in crude extracts showed a single band at pI 5.4. PCR amplification and sequencing data allowed identification of blaPER-2 and blaTEM-116. Retrospective analysis suggests that these strains were disseminated in the community, even if unnoticed, prior to their access to the hospital environment more than a decade ago.     

Prospective study ofStreptococcus milleri bacteremia

In a prospective study of bacteremia caused by organisms of theStreptococcus milleri group a total of 32 adult patients were observed over a seven-year period. These patients accounted for 1.6% of all patients diagnosed as having significant bacteremia and 17% of all cases of streptococcal (nonpneumococcal) bacteremia diagnosed during the study period. Only five patients had polymicrobic bacteremia. In 31 cases, a presumed origin of infection was identified, generally oral or gastrointestinal disease. There were only six cases of nosocomial acquisition. The most common presenting symptom was prolonged fever. The following forms of presentation were documented: bacteremia with local suppurative infection (56%), bacteremia without local suppurative infection (25%), and endocarditis (19%). An associated focus of infection was found in the abdominal cavity in 20 cases (62%). The mortality rate was 12.5%. All isolates were susceptible to penicillin. Caution is necessary in interpreting a blood culture positive forStreptococcus milleri group organisms, since, unlike other viridans streptococci, they are rarely contaminants. For this reason patients with suppurative processes and/or digestive tract disease must be carefully investigated.     

First culture isolation of Borrelia lonestari, putative agent of southern tick-associated rash illness

Southern tick-associated rash illness (STARI) is a Lyme disease-like infection described in patients in the southeastern and south-central United States, where classic Lyme disease is relatively rare. STARI develops following the bite of a lone star tick (Amblyomma americanum) and is thought to be caused by infection with an "uncultivable" spirochete tentatively named Borrelia lonestari. In this study, wild lone star ticks collected from an area where B. lonestari is endemic were cocultured in an established embryonic tick cell line (ISE6). The cultures were examined by dark-field microscopy for evidence of infection, and spirochete identity and morphology were evaluated by flagellin B and 16S rRNA gene sequence, by reaction to Borrelia-wide and B. burgdorferi-specific monoclonal antibodies, and by electron microscopy. Live spirochetes were first visualized in primary culture of A. americanum ticks by dark-field microscopy 14 days after the cell culture was inoculated. The sequences of the flagellin B and 16S rRNA genes of cultured spirochetes were consistent with previously reported sequences of B. lonestari. The cultured spirochetes reacted with a Borrelia-wide flagellin antibody, but did not react with an OspA antibody specific to B. burgdorferi, by indirect fluorescent antibody testing. Electron microscopy demonstrated organisms that were free and associated with ISE6 cells, with characteristic Borrelia sp. morphology. This study describes the first successful isolation of B. lonestari in culture, providing a much needed source of organisms for the development of diagnostic assays and forming a basis for future studies investigating the role of the organism as a human disease agent.     

Regulation of T-type calcium channels by Rho-associated kinase

We investigated the regulation of T-type channels by Rho-associated kinase (ROCK). Activation of ROCK via the endogenous ligand lysophosphatidic acid (LPA) reversibly inhibited the peak current amplitudes of rat Ca(v)3.1 and Ca(v)3.3 channels without affecting the voltage dependence of activation or inactivation, whereas Ca(v)3.2 currents showed depolarizing shifts in these parameters. LPA-induced inhibition of Ca(v)3.1 was dependent on intracellular GTP, and was antagonized by treatment with ROCK and RhoA inhibitors, LPA receptor antagonists or GDPssS. Site-directed mutagenesis of the Ca(v)3.1 alpha1 subunit revealed that the ROCK-mediated effects involve two distinct phosphorylation consensus sites in the domain II-III linker. ROCK activation by LPA reduced native T-type currents in Y79 retinoblastoma and in lateral habenular neurons, and upregulated native Ca(v)3.2 current in dorsal root ganglion neurons. Our data suggest that ROCK is an important regulator of T-type calcium channels, with potentially far-reaching implications for multiple cell functions modulated by LPA.     

Dynamic colonisation by different Pneumocystis jirovecii genotypes in cystic fibrosis patients

Although asymptomatic carriers of Pneumocystis jirovecii with cystic fibrosis (CF) have been described previously, the molecular epidemiology of P. jirovecii in CF patients has not yet been clarified. This study identified the distribution and dynamic evolution of P. jirovecii genotypes based on the mitochondrial large-subunit (mt LSU) rRNA gene. The mt LSU rRNA genotypes of P. jirovecii isolates in 33 respiratory samples from CF patients were investigated using nested PCR and direct sequencing. Three different genotypes were detected: 36.3% genotype 1 (85C/248C); 15.1% genotype 2 (85A/248C); 42.4% genotype 3 (85T/248C); and 6% mixed genotypes. Patients studied during a 1-year follow-up period showed a continuous colonisation/clearance cycle involving P. jirovecii and an accumulative tendency to be colonised with genotype 3.     

Mean cerebral blood flow measurements using phase contrast MRI in the first year of life

Alterations in cerebral blood flow (CBF) are believed to be linked to many of the neurological pathologies that affect neonates and small infants. CBF measurements are nonetheless often difficult to perform in this population, as many techniques rely on radioactive tracers or other invasive methods. In this study, mean global CBF was measured in 21 infants under the age of one, using non-invasive MRI techniques adapted to the neonatal population. Mean CBF was computed as the ratio of blood flow delivered to the brain (measured using phase contrast MRI) and brain volume (computed by segmenting anatomical MR images). Tests in adult volunteers and repeated measurements showed the flow measurements using the proposed method to be both accurate and reproducible. It was also found that cardiac gating need not be employed in infants with no known cardiac pathology. The developed technique can easily be appended to a neonatal MRI examination to provide rapid, robust, and non-invasive estimates of mean CBF, thus providing a means to monitor developmental or pathology-related alterations in cerebral perfusion and the impact of different treatment courses. In the imaged cohort, mean CBF and flow to the brain were found to rapidly increase during the first year of life (from approx. 25 to 60 ml blood/100 ml tissue/min), in good agreement with literature from other modalities where available. Mean CBF also showed a significant correlation with arterial oxygen saturation level and heart rate, but no significant correlation was found between CBF and the hematocrit or body temperature.     

Two novel AIRE mutations in autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) among Indians

Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare recessive disorder resulting from mutations in the autoimmune regulator ( AIRE ) gene. There is no information on AIRE mutations in Indians. In a cross-sectional study, nine patients (eight families), from four referral hospitals in India, were studied for AIRE mutations by direct sequencing. We screened for new mutations in 150 controls by allele-specific PCR. The patients had 1–7 known components of APECED. Three patients had unusual manifestations: presentation with type 1 diabetes; chronic sinusitis and otitis media; and facial dysmorphism. All patients carried homozygous, probably recessive, AIRE mutations. Two unrelated patients from a small in-bred community (Vanika Vaisya) in south India carried an unreported missense mutation, p.V80G, in the N-terminal caspase recruitment domain. Another unique mutation, p.C302X, resulting in a truncated protein with deletion of both zinc-finger domains, was detected in a patient from Gujarat. Neither mutation was detected in controls. Other mutations, previously described in Caucasians, were: 13 base pair deletion (p.C322fsX372) in 4 (38%), and Finn-major (p.R257X) and p.R139X (Sardinian) mutation in one subject each. In conclusion, in this first series of APECED in Indians, we detected AIRE mutations previously reported in Caucasians, as well as unique mutations. Of these, p.V80G is possibly an ancestral mutation in an in-bred community.     

Epidemiology of extended-spectrum beta-lactamase-producing Enterobacter isolates in a Spanish hospital during a 12-year period

Fifteen Enterobacter clinical isolates (11 Enterobacter cloacae isolates, 3 Enterobacter aerogenes isolates, and 1 Enterobacter gergoviae isolate), representing 0.4% of all Enterobacter isolates recovered in our hospital from 1989 to 2000, were suspected of harboring an extended-spectrum beta-lactamase (ESBL). These isolates were recovered from 14 different patients. ESBLs were transferred by conjugation into an Escherichia coli recipient strain. Pulsed-field gel electrophoresis (PFGE) revealed a single clone of E. aerogenes and six different clones of E. cloacae. Four of these E. cloacae clonal types were represented by only one isolate each, but the other two were represented by three and four isolates, respectively. Isoelectric focusing, susceptibility phenotyping, PCR analysis, and sequencing demonstrated the presence of three different ESBLs. The most frequent was the recently characterized CTX-M-10 ESBL, which was found in the E. gergoviae isolate and in all but one of the E. cloacae isolates. The remaining E. cloacae isolate harbored a TEM-27 ESBL, and the three E. aerogenes isolates harbored a TEM-24 ESBL. PFGE revealed that our E. aerogenes strain was indistinguishable from the French TEM-24-producing E. aerogenes endemic clone. Although a low prevalence of ESBL-producing Enterobacter isolates was found in our institution over a 12-year period, a diversity of nonepidemic E. cloacae clones was detected, as was the persistence of the CTX-M-10 beta-lactamase. The presence of the TEM-24-producing E. aerogenes French clone in our institution also demonstrates the intercountry dissemination of ESBL-producing isolates.