Respiratory viruses in the pediatric intensive care unit: prevalence and clinical aspects

A survey was conducted in two pediatric intensive care units in hospitals in Porto Alegre, Brazil, in order to monitor the main respiratory viruses present in bronchiolitis and/or pneumonia and their involvement in the severity of viral respiratory infections. Viral respiratory infection prevalence was 38.7%. In bronchiolitis, respiratory syncytial virus (RSV) was detected in 36% of the cases. In pneumonia, the prevalence rates were similar for adenovirus (10.3%) and RSV (7.7%). There was a difference among the viruses detected in terms of frequency of clinical findings indicating greater severity. Frequency of crackles in patients with RSV (47.3%) showed a borderline significance (p = 0.055, Fisher's exact test) as compared to those with adenovirus (87.5%). The overall case fatality rate in this study was 2.7%, and adenovirus showed a significantly higher case fatality rate (25%) than RSV (2.8%) (p = 0.005). Injected antibiotics were used in 49% of the children with RSV and 60% of those with adenovirus. Adenovirus was not detected in any of the 33 children submitted to oxygen therapy.

The functional outcome and recovery of patients admitted to an intensive care unit following drug overdose: a follow-up study.

Patients who have overdosed on drugs commonly present to emergency departments, with only the most severe cases requiring intensive care unit (ICU) admission. Such patients typically survive hospitalisation. We studied their longer term functional outcomes and recovery patterns which have not been well described. All patients admitted to the 18-bed ICU of a university-affiliated teaching hospital following drug overdoses between 1 January 2004 and 31 December 2006 were identified. With ethical approval, we evaluated the functional outcome and recovery patterns of the surviving patients 31 months after presentation, by telephone or personal interview. These were recorded as Glasgow outcome score, Karnofsky performance index and present work status. During the three years studied, 43 patients were identified as being admitted to our ICU because of an overdose. The average age was 34 years, 72% were male and the mean APACHE II score was 16.7. Of these, 32 were discharged from hospital alive. Follow-up data was attained on all of them. At a median of 31 months follow-up, a further eight had died. Of the 24 surviving there were 13 unemployed, seven employed and four in custody. The median Glasgow outcome score of survivors was 4.5, their Karnofsky score 80. Admission to ICU for treatment of overdose is associated with a very high risk of death in both the short- and long-term. While excellent functional recovery is achievable, 16% of survivors were held in custody and 54% unemployed.This resource was contributed by The National Documentation Centre on Drug Use.

Daisy Roots Garden Project

In the summer of 2009 a local Men’s group that met in St Helena’s thought that it might be a good idea to sow a wild garden on the site.  Then came the raised beds – 2 were placed in the garden area opposite the front door of the house and as it was winter there was not much happening and they were quickly renamed the ‘coffins’.  This was the start of ‘Daisy Roots’. Dublin City Council provided top soil and water butts.  An Taisce funded the start up phase of the project and now continue to provide funds for insurance and some development work.  At the right hand side of St Helena’s there is, what was the old stable yard of the ‘big house’.  This had been used to house a large ‘lock up container and a porta-cabin.  These have been now removed and this is where the main part of the garden is. Following a conference of Business in the Community Ireland, DX Ltd expressed an interest in getting involved in a local project.  Together DX staff and the volunteers reclaimed the old stables and gave them new roofs and new doors.  Paths were put in between the raised vegetable beds and new raised beds were built.  A seating area was developed.  The garden is now wheelchair accessible and there are wheelchair accessible flowerbeds in the garden also.  The newest projects in the garden are a memorial flowerbed and a propagator. Dublin City Council via Cherry Orchard Regeneration Board Initiative Type Community Food Growing Projects Location Dublin 11 Funding Dublin City Council via Cherry Orchard Regeneration Board Partner Agencies An Taisce Dublin City Council HSE Tolka Area Partnership

Fettercairn Growing Community Roots

Fettercairn Growing Community Roots will deliver a range of programmes around exercise and healthy eating to improve, encourage and integrate the target schools into the wider local community through linkages with community based projects e.g. Fettercairn Community Garden. Part of theCFI Programme 2013-2015 Initiative Type Community Food Growing Projects Nutrition Education and Training Programmes Location Dublin 24 Partner Agencies safefood

Candida colonization in intensive care unit patients' urine

The objective of this study was to identify possible predisposing factors for candiduria in intensive care unit (ICU) patients from Hospital das Clínicas, Universidade Federal de Goiás, Goiânia, Brazil, during one year. Urine samples from 153 ICU patients were obtained by catheterization on admission day and every seven days. Data such as sex, age, antifungal therapy, and variables as antibiotics, underlying diseases or comorbid conditions and stay in the hospital, were collected from patients who had at least one urine culture that yielded > 10³ yeast colonies/ml. Candiduria was recovered in 68 patients and the commonest predisposing factors were antibiotic therapy (100%) and indwelling urinary catheter (92.6%). The percentage of Candida spp. isolation increased during the extended periods in which patients remained in the ICU. C. albicans was isolated in 69.1%, and the other species non-albicans as C. glabrata, C. kefyr, C. parapsilosis, C. famata, C. guilliermondii, C. krusei, and C. tropicalis were isolated in lower percentage. The high frequency of candiduria and the possible predisposing factors found in ICU patients show that candiduria surveillance should be performed to help reducing nosocomial infections.

U-Pb ages of magmatic rocks of the western Austroalpine Dent-Blanche-Sesia unit

Conventional U-Pb ages on zircon and monazite demonstrate that granites and gabbros intruded during a short time span of 5 Ma between 293 and 288 Ma in several polycyclic basement units of the Western Austroalpine domain. This bimodal activity reflects increasing underplating of an upwelling mantle at the base of a thinning post-Variscan continental crust.

Trypanosoma cruzi strains isolated from human, vector, and animal reservoir in the same endemic region in Mexico and typed as T. cruzi I, discrete typing unit 1 exhibit considerable biological diversity

In this study, three strains of Trypanosoma cruzi were isolated at the same time and in the same endemic region in Mexico from a human patient with chronic chagasic cardiomyopathy (RyC-H); vector (Triatoma barberi) (RyC-V); and rodent reservoir (Peromyscus peromyscus) (RyC-R). The three strains were characterized by multilocus enzyme electrophoresis, random amplified polymorphic DNA, and by pathological profiles in experimental animals (biodemes). Based on the analysis of genetic markers the three parasite strains were typed as belonging to T. cruzi I major group, discrete typing unit 1. The pathological profile of RyC-H and RyC-V strains indicated medium virulence and low mortality and, accordingly, the strains should be considered as belonging to biodeme Type III. On the other hand, the parasites from RyC-R strain induced more severe inflammatory processes and high mortality (> 40%) and were considered as belonging to biodeme Type II. The relationship between genotypes and biological characteristics in T. cruzi strains is still debated and not clearly understood. An expert committee recommended in 1999 that Biodeme Type III would correspond to T. cruzi I group, whereas Biodeme Type II, to T. cruzi II group. Our findings suggest that, at least for Mexican isolates, this correlation does not stand and that biological characteristics such as pathogenicity and virulence could be determined by factors different from those identified in the genotypic characterization

Re-mapping the molecular features of the human immunodeficiency virus type 1 and human T-cell lymphotropic virus type 1 Brazilian sequences using a bioinformatics unit established in Salvador, Bahia, Brazil, to give support to the viral epidemiology studies

The analysis of genetic data for human immunodeficiency virus type 1 (HIV-1) and human T-cell lymphotropic virus type 1 (HTLV-1) is essential to improve treatment and public health strategies as well as to select strains for vaccine programs. However, the analysis of large quantities of genetic data requires collaborative efforts in bioinformatics, computer biology, molecular biology, evolution, and medical science. The objective of this study was to review and improve the molecular epidemiology of HIV-1 and HTLV-1 viruses isolated in Brazil using bioinformatic tools available in the Laboratório Avançado de Sáude Pública (Lasp) bioinformatics unit. The analysis of HIV-1 isolates confirmed a heterogeneous distribution of the viral genotypes circulating in the country. The Brazilian HIV-1 epidemic is characterized by the presence of multiple subtypes (B, F1, C) and B/F1 recombinant virus while, on the other hand, most of the HTLV-1 sequences were classified as Transcontinental subgroup of the Cosmopolitan subtype. Despite the high variation among HIV-1 subtypes, protein glycosylation and phosphorylation domains were conserved in the pol, gag, and env genes of the Brazilian HIV-1 strains suggesting constraints in the HIV-1 evolution process. As expected, the functional protein sites were highly conservative in the HTLV-1 env gene sequences. Furthermore, the presence of these functional sites in HIV-1 and HTLV-1 strains could help in the development of vaccines that pre-empt the viral escape process.

Use of the pyrophosphatase activity as a reliable tonoplast marker in maize roots.

Membranes of maize (Zea mays L., cv LG 11) roots were fractionated by sucrose (in presence or absence of Mg2+) or dextran density gradient centrifugations and the locations of organelles were determined using marker enzymes. Latent UDPase was used as a Golgi marker, catalase for the peroxysomes, cytochrome c oxidase for the mitochondria, UDP-Gal-galactosyltransferase for the amyloplast membranes and NADH-cytochrome c reductase for the ER. Two markers were selected for the plasmalemma, the vanadate-sensitive ATPase and UDP-Glc-sterolglucosyltransferase. The distributions of the PPase and vacuolar ATPase were found to be similar after density gradient centrifugation. The PPase and vacuolar ATPase activities were clearly separated from almost all the other markers tested, however, a partial association of both activities with the ER cannot be completely ruled out. The PPase of maize roots is more active and easier to measure than the vacuolar ATPase and is therefore an excellent candidate for use as a tonoplast marker.

A Ca2+/H+ antiport system driven by the tonoplast pyrophosphate-dependent proton pump from maize roots.

Calcium uptake by tonoplast enriched membrane vesicles from maize (Zea mays L. cv. LG 11) primary roots was studied. A pH gradient, measured by the fluorescence quenching of quinacrine, was generated across sealed vesicles driven by the pyrophosphate-dependent proton pump. The fluorescence quenching was strongly inhibited by Ca2+; moreover, when increasing Ca2+ concentrations were added to vesicles at steady-state, a concomitant decrease in the proton gradient was observed. Ca2+ uptake using Ca-45(2+) was linear from 10 min when oxalate (10 mM) was present, while Ca2+ uptake was completely inhibited with proton ionophores (FCCP and monensin), indicating a Ca2+/H+ antiport. Membranes were further fractionated using a linear sucrose density gradient (10-45%) and were identified with marker enzymes. Ca2+ uptake co-migrated with the tonoplast pyrophosphate-dependent proton pumping, pyrophosphatase and ATPase activities: the Ca2+/H+ antiport is consequently located at the tonoplast.

Tonoplast localization of a calmodulin-stimulated Ca2+-pump from maize roots.

The subcellular localization of a calmodulin-stimulated calcium (Ca2+)-ATPase activity from maize roots (Zea mays L., cv LG 11) was studied. For this purpose, an efficient procedure was developed to prepare sealed plasma membrane vesicles allowing the measurement of proton and Ca2+ transport activities. Two-day-old root membranes were fractionated by sucrose and dextran density gradient centrifugation. Marker enzymes were used to study the distribution of the different membranes in the gradients and a filtration technique was developed to measure Ca-45(2+) transport in sealed vesicles. Most of the ATP-dependent Ca2+ transport activity was associated with the ER. However, a small part of this activity was associated with the tonoplast (corresponding to the activity of the H+/Ca2+ antiport) and the plasma membrane. When the Ca2+ transport was measured in the presence of exogenous calmodulin (1 muM), a 3-5-fold increase of uptake was measured. The calmodulin-stimulated activity was associated with the tonoplast vesicles only. This activity was insensitive to monensin, a proton ionophore, ruling out a direct effect of calmodulin on the H+/Ca2+ antiport. In conclusion, four different Ca2+ transporters are present in young maize root cells. A Ca2+/H+ antiport system is present on the tonoplast, whereas, the plasma membrane and the ER possess each a calmodulinin-sensitive Ca2+-ATPase. Finally, a calmodulin-stimulated Ca2+-ATPase is associated with the tonoplast.

Characterization of the tonoplast Ca2+/H+ antiport system from maize roots.

The tonoplast calcium Ca2+/H+ antiport system of maize (Zea mays L. cv LG 11) roots was characterized using the ''pH jump'' technique in order to avoid interference from the tonoplast proton and Ca2+ pumps. Ca2+ uptake was recorded in the presence of different inhibitors and divalent ions. Chemical modification of amino acid residues of the antiport was used to elucidate the amino acid residues participating in the Ca2+ transport activity. The Ca2+/H+ antiport activity was found to be strongly inhibited by ruthenium red and verapamil, whereas diethylstilbestrol was less effective. Vanadate, erythrosin B, cyclopiazonic acid, bafilomycin, thapsigargin, N,N'-dicyclohexylcarbodiimide (DCCD) and 4,4'-diisothiocyanatostilbene-2,2'-disulfonate (DIDS) were without effect. Lanthanum and divalent ions were strongly inhibitory (Cd2+ > Mn2+ > Sr2+ > Ba2+). While reagents modifying sulfhydryl groups (N-ethylmaleimide and 5,5'-dithio-bis(2-nitrobenzoate)) did not affect the antiport activity, modification of trytophan residues (N-bromosuccinimide) was strongly inhibitory. We conclude that ruthenium red, verapamil, lanthanum and divalent cations directly inhibit Ca2+ uptake independent of the function of the proton and Ca2+ pumps. Moreover, the results of chemically modified amino acid residues suggest that sulfhydryl groups are not involved in Ca2+ transport, while tryptophan residues seem important for this translocation.

Subcellular localization, distribution and expression of calmodulin in Zea mays roots.

The subcellular localization, distribution and the steady state level of calmodulin from maize roots (Zea mays L., cv. LG 11) were studied. To analyze the subcellular localization, 2-day old root membranes were fractionated by sucrose density gradient centrifugation and immunoblotting was done with an antibody raised against a vertebrate calmodulin (SWant) which recognized the plant calmodulin. Calmodulin was principally associated with high density fractions and particularly plasmalemma. For studying the distribution of calmodulin in various zones of Zea mays roots, a micro method of membrane preparation was developed. Most of the calmodulin was present in microsomes isolated from the root apex corresponding to the first 4 mm of a 15 +/- 2 mm root. An identical distribution was found by studying the steady state level of the protein by Northern blotting using a cDNA clone of Zea mays calmodulin.