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Amyloidosis is a generic term that refers to the deposition of amyloid fibrils in bodily tissues. Its onset is usually after 40 years of age, with localized or systemic involvement associated with multiple myeloma or chronic inflammatory diseases, and can mimic various rheumatic syndromes. We report the case of a patient with amyloidosis associated with multiple myeloma, showing clinical characteristics of pseudomyopathy.

Objective: To assess the association of demographic and clinical aspects with radiographically diagnosed pneumonia., Design: By active surveillance, the admitted pneumonia cases by the pediatrician on duty were identified in a 2 year period. Demographic, clinical and radiographic data were registered into standardized forms., Setting: A public university pediatric hospital in Salvador, Northeast Brazil., Patients: Children <5 years-old., Main Outcome Measures: Radiographically diagnosed pneumonia based on detection of pulmonary infiltrate/ consolidation., Results: 301 cases had the chest X-ray evaluated by a pediatric radiologist blinded to clinical information, among whom pulmonary infiltrate and consolidation were described in 161 (54%) and 119 (40%), respectively. Chest X-ray was read normal for 140 cases. Overall, the median age was 17 months (mean 20±14, range 12 days-59 months). Pulmonary infiltrate was less frequently described among patients aged under 1 year (41.3% vs 59.9%, P=0.002, OR [95% CI] = 0.47 [0.29-0.76]) and hyperinflation was significantly more frequent in this age group (27.9% vs 4.1%, P<0.001, OR [95% CI] = 9.14 [4.0-20.9]). By multiple logistic regression, fever on admission was independently associated with pulmonary infiltrate (OR [95% CI] = 1.68 [1.03-2.73]) or consolidation (1.79 [1.10-2.92]), wheezing was independently associated with absence of pulmonary infiltrate (0.53 [0.33-0.86]) or of consolidation (0.53 [0.33-0.87]). The positive likelihood ratio of fever on examination for pulmonary infiltrate and consolidation was 1.49 (95% CI:1.11-1.98) and 1.49 (95% CI: 1.14-1.94), respectively., Conclusion: Presence of fever enhanced 2.5 times the chance of children hospitalized with lower respiratory tract disease to have radiographically diagnosed pneumonia.

(Bacterio)phage PVP-SE1, isolated from a German wastewater plant, presents a high potential value as a biocontrol agent and as a diagnostic tool, even compared to the well-studied typing phage Felix 01, due to its broad lytic spectrum against different Salmonella strains. Sequence analysis of its genome (145,964 bp) shows it to be terminally redundant and circularly permuted. Its G+C content, 45.6 mol%, is lower than that of its hosts (50 to 54 mol%). We found a total of 244 open reading frames (ORFs), representing 91.6% of the coding capacity of the genome. Approximately 46% of encoded proteins are unique to this phage, and 22.1% of the proteins could be functionally assigned. This myovirus encodes a large number of tRNAs (n=24), reflecting its lytic capacity and evolution through different hosts. Tandem mass spectrometric analysis using electron spray ionization revealed 25 structural proteins as part of the mature phage particle. The genome sequence was found to share homology with 140 proteins of the Escherichia coli bacteriophage rV5. Both phages are unrelated to any other known virus, which suggests that an "rV5-like virus" genus should be created within the Myoviridae to contain these two phages.

The ICNP® model of seven axes resulted in the transfer of terms between the axes, conceptual modifications, inclusion and exclusion of terms. This descriptive, bibliographic study, mapped out the terms of the axes "Time", "Location", "Means" and "Client" among Beta-2, 1.0 and 1.1 versions of the ICNP® and CIPESC®. The mapping located the terms of these axes in the different versions of the ICNP® and CIPESC®. It was performed to classify the semantic equivalence of terms and concepts. The result shows that some changes have the objective of reducing ambiguities and redundancies; others do not allow the verification of the criterion for exclusion, inclusion and migration of terms. It is imperative to establish concepts of consensus by the group responsible for building the terminology in order to minimize the misuse or improper use of terms. It is necessary to revise the list of actions in the vocabulary inventory of CIPESC®.

We identified complex genomic rearrangements consisting of intermixed duplications and triplications of genomic segments at the MECP2 and PLP1 loci. These complex rearrangements were characterized by a triplicated segment embedded within a duplication in 11 unrelated subjects. Notably, only two breakpoint junctions were generated during each rearrangement formation. All the complex rearrangement products share a common genomic organization, duplication-inverted triplication-duplication (DUP-TRP/INV-DUP), in which the triplicated segment is inverted and located between directly oriented duplicated genomic segments. We provide evidence that the DUP-TRP/INV-DUP structures are mediated by inverted repeats that can be separated by >300 kb, a genomic architecture that apparently leads to susceptibility to such complex rearrangements. A similar inverted repeat-mediated mechanism may underlie structural variation in many other regions of the human genome. We propose a mechanism that involves both homology-driven events, via inverted repeats, and microhomologous or nonhomologous events.

Photodynamic therapy is a very promising approach to inactivate pathogenic microorganisms. The photodamage of cells involves reactive oxygen species (ROS) which are generated in situ by two main mechanisms (type I and/or type II). The mechanism responsible for the photoinactivation (PI) of a bioluminescent recombinant Escherichia coli, induced by three different cationic porphyrins, was identified in this work using a rapid method based on the monitoring of the metabolic activity of this bacterium. The inhibitory effect of the photodynamic process in the presence of a singlet oxygen quencher (sodium azide) or free radical scavengers (d-mannitol and l-cysteine) was evaluated by exposing bacterial suspensions with 0.5 μM Tri-Py(+)-Me-PF, 5.0 μM Tetra-Py(+)-Me or 5.0 μM Tri-SPy(+)-Me-PF to white light. Strong bacterial protection was observed with sodium azide (100 mM) for the three cationic porphyrins. However, in the presence of Tri-Py(+)-Me-PF and Tetra-Py(+)-Me and the free radical scavengers (l-cysteine and d-mannitol) the reduction on the bacterial bioluminescence was significantly higher and similar to that obtained in their absence (5.4-6.0 log reduction). In the case of Tri-SPy(+)-Me-PF two distinct behaviours were observed when l-cysteine and d-mannitol were used as free radical scavengers: while the presence of l-cysteine (100 mM) lead to a bacterial protection similar to the one observed with sodium azide, in the presence of d-mannitol only a small protection was detected. The high inhibition of the PS activity by l-cysteine is not due to its radical scavenger ability but due to the singlet oxygen quenching by the sulfanyl group (-SH). In fact, the photodecomposition of 1,3-diphenylisobenzofuran in the presence of Tri-SPy(+)-Me-PF is completely suppressed when l-cysteine is present. The results obtained in this study suggest that singlet oxygen (type II mechanism) plays a very important role over free radicals (type I mechanism) on the PI process of the bioluminescent E. coli by Tri-Py(+)-Me-PF, Tetra-Py(+)-Me and Tri-SPy(+)-Me-PF. Although the use of scavengers is an adequate and simple approach to evaluate the relative importance of the two pathways, it is important to choose scavengers which do not interfere in both PI mechanisms. Sodium azide and d-mannitol seem to be good oxygen and free radical quenchers, respectively, to study the PI mechanisms by porphyrinic photosensitizers., (This journal is © The Royal Society of Chemistry and Owner Societies 2011)

Background: Limited information is available on the role of human metapneumovirus (HMPV) as the unique pathogen among children hospitalized for community-acquired pneumonia (CAP) in a tropical region., Objective: We aimed to describe HMPV infection among children with CAP investigating bacterial and viral co-infections., Patients and Methods: A prospective study was carried out in Salvador, North-East Brazil. Overall, 268 children aged <5 years hospitalized for CAP were enrolled. Human metapneumovirus RNA was detected in nasopharyngeal aspirates (NPA) by reverse transcription polymerase chain reaction. Sixteen other bacterial and viral pathogens were investigated by an expanded panel of laboratory methods. Chest X-ray taken on admission was read by an independent paediatric radiologist unaware of clinical information or the established aetiology., Results: Human metapneumovirus RNA was detected in NPAs of 11 (4.1%) children, of which 4 (36%) had sole HMPV infection. The disease was significantly shorter among patients with sole HMPV infection in comparison with patients with mixed infection (4 ± 1 versus 7 ± 2 days, P = 0.03). Three of those four patients had alveolar infiltrates., Conclusion: Sole HMPV infection was detected in children with CAP in Salvador, North-East Brazil. HMPV may play a role in the childhood CAP burden., (© 2011 Blackwell Publishing Ltd.)

Purpose: Autosomal dominant spastic paraplegia, type 4 (SPG4), a debilitating disorder of progressive spasticity and weakness of the lower limbs, results from heterozygous mutations in the SPAST gene. The full spectrum of SPAST mutations causing SPG4 and their mechanisms of formation remain to be determined., Methods: We used multiplex ligation-dependent probe amplification, locus-specific array comparative genomic hybridization, and breakpoint DNA sequencing to identify and describe genomic rearrangements in three patients with a clinical presentation of hereditary spastic paraplegia., Results: We describe three SPG4 patients with intragenic rearrangements in SPAST; all specifically delete the final exon, exon 17. Breakpoint sequence analyses provide evidence for Alu-specific microhomology-mediated deletion as the mechanism of exon loss; one complex rearrangement apparently occurred by multiple Alu-facilitated template switches., Conclusion: We hypothesize that the high concentration of Alu family members in the introns and flanking sequence of SPAST may predispose to intragenic rearrangements. Thus, Alu-specific microhomology-mediated intragenic rearrangements in SPAST may be a common cause of SPG4. Furthermore, we propose that genomic deletions encompassing the final exon of SPAST may affect expression of SLC30A6, the most proximal downstream locus and a gene that has been implicated in the pathogenesis of Alzheimer disease, potentially explaining recent reports of dementia in selected SPG4 patients.

Genomic instability is a feature of the human Xp22.31 region wherein deletions are associated with X-linked ichthyosis, mental retardation and attention deficit hyperactivity disorder. A putative homologous recombination hotspot motif is enriched in low copy repeats that mediate recurrent deletion at this locus. To date, few efforts have focused on copy number gain at Xp22.31. However, clinical testing revealed a high incidence of duplication of Xp22.31 in subjects ascertained and referred with neurobehavioral phenotypes. We systematically studied 61 unrelated subjects with rearrangements revealing gain in copy number, using multiple molecular assays. We detected not only the anticipated recurrent and simple nonrecurrent duplications, but also unexpectedly identified recurrent triplications and other complex rearrangements. Breakpoint analyses enabled us to surmise the mechanisms for many of these rearrangements. The clinical significance of the recurrent duplications and triplications were assessed using different approaches. We cannot find any evidence to support pathogenicity of the Xp22.31 duplication. However, our data suggest that the Xp22.31 duplication may serve as a risk factor for abnormal phenotypes. Our findings highlight the need for more robust Xp22.31 triplication detection in that such further gain may be more penetrant than the duplications. Our findings reveal the distribution of different mechanisms for genomic duplication rearrangements at a given locus, and provide insights into aspects of strand exchange events between paralogous sequences in the human genome.

Aim: The aim of this study was to evaluate, by light microscopy, the effects of laser phototherapy (LPT) at 780 nm or a combination of 660 and 790 nm, on the inflammatory process of the rat temporomandibular joint (TMJ) induced by carrageen., Background: Temporomandibular disorders (TMDs) are frequent in the population and generally present an inflammatory component. Previous studies have evidenced positive effects of laser phototherapy on TMDs. However, its mechanism of action on the inflammation of the TMJ is not known yet., Materials and Methods: Eighty-five Wistar rats were divided into 9 groups: G1, Saline; G2, Saline + LPT IR; G3, Saline + LPT IR + R; G4, Carrageenan; G5, Carrageenan + LPT IR; G6, Carrageenan + LPT IR + R; G7, previous LPT + Carrageenan; G8, previous LPT + carrageenan + LPT IR; and G9, previous LPT + carrageenan + LPT IR + R, and then subdivided in subgroups of 3 and 7 days. After animal death, specimens were taken, routinely cut and stained with HE, Sirius Red, and Toluidine Blue. Descriptive analysis of components of the TMJ was done. The synovial cell layers were counted., Results: Injection of saline did not produced inflammatory reaction and the irradiated groups did not present differences compared to nonirradiated ones. After carrageenan injection, intense inflammatory infiltration and synovial cell layers proliferation were observed. The infrared irradiated group presented less inflammation and less synovial cell layers number compared to other groups. Previous laser irradiation did not improve the results., Conclusion: It was concluded that the LPT presented positive effects on inflammatory infiltration reduction and accelerated the inflammation process, mainly with IR laser irradiation. The number of synovial cell layers was reduced on irradiated group.

Documental descriptive research, presents the mapping of the terms from the "Action" Axis among the versions: Beta-2, 1.0 and 1.1 of the ICNP® and the verbs of CIPESC®. Results are: 22% of terms from Version 1.0 are new; 12.61% are identical, with different concept; 51% are not in CIPESC®; 65% of terms from CIPESC® are not in ICNP®, in relation to them, 15% represent actions related to link building or empowerment. The results require validation by experts. This paper discusses the complexity of the Portuguese language and the need to establish consensus among concepts. The lack of verbs from CIPESC® is considered a gap to be filled, which may reflect the use of ICNP® by nurses in public health.

Genomic disorders are defined as diseases caused by rearrangements of the genome incited by a genomic architecture that conveys instability. Y-chromosome related dysfunctions such as male infertility are frequently associated with gross DNA rearrangements resulting from its peculiar genomic architecture. The Y-chromosome has evolved into a highly specialized chromosome to perform male functions, mainly spermatogenesis. Direct and inverted repeats, some of them palindromes with highly identical nucleotide sequences that can form DNA cruciform structures, characterize the genomic structure of the Y-chromosome long arm. Some particular Y chromosome genomic deletions can cause spermatogenic failure likely because of removal of one or more transcriptional units with a potential role in spermatogenesis. We describe mechanisms underlying the formation of human genomic rearrangements on autosomes and review Y-chromosome deletions associated with male infertility.

Objectives: To evaluate the effects of acupuncture and sham-acupuncture on women with menopausal symptoms as reflected in the intensity of their hot flushes and the Kupperman Menopausal Index (KMI)., Method: This was a randomized, single-blind, placebo-controlled, cross-over trial with 81 patients assigned to two groups: Group 1 received 12 months of acupuncture, then 6 months of sham-acupuncture treatment (n = 56) and Group 2 received 6 months of sham-acupuncture, then 12 months of acupuncture treatment (n = 25). The needles were inserted in a harmonic craniocaudal manner at a depth of about 2 cm, and each session lasted approximately 40  min. The efficacy of acupuncture in ameliorating the climacteric symptoms of patients in postmenopause was determined through the KMI and the intensity of hot flushes. The analysis of variance method for two factors and repeated measures was applied., Results: The baseline values of the women in both groups were similar for the KMI score and number of hot flushes. At the end of 6 months, the values for the KMI and hot flushes for the women in Group 1 were lower than those of the women in Group 2 (p < 0.05). After 12 months, the KMI and hot flush data were similar in both groups. After 18 months, the values of the KMI and hot flushes for the women in Group 2 for were lower than those of the women in Group 1 (p < 0.05)., Conclusion: Acupuncture treatment for relieving menopausal symptoms may be effective for decreasing hot flushes and the KMI score in postmenopausal women.

A novel 1,3-dioxopyrrolo[3,4-b]porphyrin (2) has been synthesized in 70% yield following a [4 + 2] cycloaddition reaction of pyrrolo[3,4-b]porphyrin 1 with (1)O(2). The new imide was used as a template to other 1,3-dioxopyrrolo[3,4-b]porphyrins and to the corresponding open counterparts. The UV/vis absorption spectra of the new compounds show significant red-shifts when compared with those of the nonsubstituted analogues. The structure of an imide derivative was confirmed by single-crystal X-ray diffraction.

Introduction: Advanced breast cancer is a very frequent serious public health problem., Objective: This study aimed to analyze the prevalence of arm symptoms and overall survival in affected patients., Methodology: A retrospective cohort study was accomplished in 196 women presenting with advanced breast cancer. A descriptive analysis was carried out by and overall survival probabilities were calculated according to Kaplan-Meier method and Cox regression., Results: These included 196 woman had a medium age of 55.9 years, the majority being obese (71.3%) and having a IIIB clinical staging (52.3%). The most prevalent arm symptoms were pain (54.6%), paresthesia in upper limb (8.5%), lymphedema (> 200 ml) (6.6%), shoulder restriction (4.7%) and axillary web syndrome (0.6%). The median follow-up was 18.7 months (SD 7,33), 4.0% of the patients had metastasis and 31.6% were dead. The mean free disease survival was 20.4 months (95% CI 18.4-22.4), and the mean survival time was 26.3 months (95% CI 24.7-28.0). After adjusted the association with arm symptom and overall survival for clinical stage and presence of distant metastasis, the shoulder restriction was a predict variable, were represents the risk 2.59 for worse survival in compared with woman with no shoulder restriction (CI 95% 1.06-5.98)., Conclusion: The population studied at this study presented high prevalence of complications earlier than neo adjuvant treatment. After controlling for effects of tumor variables, the shoulder restriction was statistically associated with poor overall survival after 2 years.

Objective: To determine prevalence of overweight and abdominal fat in adult population in the urban area of Teresina-PI., Methods: Cross-sectional study with probabilistic sample by conglomerates. The study evaluated 464 adults, 20 to 59 years of age living in the urban area of Teresina-PI. Nutritional status was classified by the body mass index (BMI) and abdominal fat accumulation was estimated according to waist circumference. The significance level was set at 5% (p<0.05)., Results: Prevalence of overweight and obesity according to nutritional status based on BMI was, respectively, 30.0% and 7.7%. There was an increase in the proportion of overweight and obesity among men with progressively higher family income. Highest rates of obesity were found among individuals 50 to 59 years of age with stable marriages and nonsmokers. No association was found between individual or family income and presence of abdominal fat in the population., Conclusion: Prevalence of overweight in the study population follows the Brazilian trend. Proportions of overweight and obesity were higher among men and increased with age. Women and married persons showed a greater tendency for abdominal obesity.

Objective: To describe the evolution and outcome of children hospitalized with community-acquired pneumonia receiving penicillin., Methods: A search was carried out for all hospitalized community-acquired pneumonia cases in a 37-month period. Inclusion criteria comprised age >2 months, intravenous penicillin G use at 200,000 IU/kg/day for >48 h and chest x-ray results. Confounders leading to exclusion included underlying debilitating or chronic pulmonary illnesses, nosocomial pneumonia or transference to another hospital. Pneumonia was confirmed if a pulmonary infiltrate or pleural effusion was described by an independent radiologist blind to the clinical information. Data on admission and evolution were entered on a standardized form., Results: Of 154 studied cases, 123 (80%) and 40 (26%) had pulmonary infiltrate or pleural effusion, respectively. Penicilli was substituted by other antibiotics in 28 (18%) patients, in whom the sole significant decrease was in the frequency of tachypnea from the first to the second day of treatment (86% vs. 50%, p = 0.008). Among patients treated exclusively with penicillin G, fever (46% vs. 26%, p = 0.002), tachypnea (74% vs. 59%, p = 0.003), chest indrawing (29% vs. 13%, p<0.001) and nasal flaring (10% vs. 1.6%, p = 0.001) frequencies significantly decreased from admission to the first day of treatment. Patients treated with other antimicrobial agents stayed longer in the hospital than those treated solely with penicillin G (16 ± 6 vs. 8 ± 4 days, p<0.001, mean difference (95% confidence interval) 8 (6-10)). None of the studied patients died., Conclusion: Penicillin G successfully treated 82% (126/154) of the study group and improvement was marked on the first day of treatment.

Mercury toxicity is a highly interesting topic in biomedicine due to the severe endpoints and treatment limitations. Selenite serves as an antagonist of mercury toxicity, but the molecular mechanism of detoxification is not clear. Inhibition of the selenoenzyme thioredoxin reductase (TrxR) is a suggested mechanism of toxicity. Here, we demonstrated enhanced inhibition of activity by inorganic and organic mercury compounds in NADPH-reduced TrxR, consistent with binding of mercury also to the active site selenolthiol. On treatment with 5 μM selenite and NADPH, TrxR inactivated by HgCl(2) displayed almost full recovery of activity. Structural analysis indicated that mercury was complexed with TrxR, but enzyme-generated selenide removed mercury as mercury selenide, regenerating the active site selenocysteine and cysteine residues required for activity. The antagonistic effects on TrxR inhibition were extended to endogenous antioxidants, such as GSH, and clinically used exogenous chelating agents BAL, DMPS, DMSA, and α-lipoic acid. Consistent with the in vitro results, recovery of TrxR activity and cell viability by selenite was observed in HgCl(2)-treated HEK 293 cells. These results stress the role of TrxR as a target of mercurials and provide the mechanism of selenite as a detoxification agent for mercury poisoning.

Objective: To examine the ability of the criteria proposed by the WHO to identify pneumonia among cases presenting with wheezing and the extent to which adding fever to the criteria alters their performance., Design: Prospective classification of 390 children aged 2-59 months with lower respiratory tract disease into five diagnostic categories, including pneumonia. WHO criteria for the identification of pneumonia and a set of such criteria modified by adding fever were compared with radiographically diagnosed pneumonia as the gold standard., Results: The sensitivity of the WHO criteria was 94% for children aged <24 months and 62% for those aged ≥24 months. The corresponding specificities were 20% and 16%. Adding fever to the WHO criteria improved specificity substantially (to 44% and 50%, respectively). The specificity of the WHO criteria was poor for children with wheezing (12%). Adding fever improved this substantially (to 42%). The addition of fever to the criteria apparently reduced their sensitivity only marginally (to 92% and 57%, respectively, in the two age groups)., Conclusion: The authors' results reaffirm that the current WHO criteria can detect pneumonia with high sensitivity, particularly among younger children. They present evidence that the ability of these criteria to distinguish between children with pneumonia and those with wheezing diseases might be greatly enhanced by the addition of fever.

Cationic nanomagnet-porphyrin hybrids were synthesized and their photodynamic therapy capabilities were investigated against the Gram (-) Escherichia coli bacteria, the Gram (+) Enterococcus faecalis bacteria and T4-like phage. The synthesis, structural characterization, photophysical properties, and antimicrobial activity of these new materials are discussed. The results show that these new multicharged nanomagnet-porphyrin hybrids are very stable in water and highly effective in the photoinactivation of bacteria and phages. Their remarkable antimicrobial activity, associated with their easy recovery, just by applying a magnetic field, makes these materials novel photosensitizers for water or wastewater disinfection.

Community-acquired pneumonia (CAP) is a common cause of morbidity among children. Evidence on seasonality, especially on the frequency of viral and bacterial causative agents is scarce; such information may be useful in an era of changing climate conditions worldwide. To analyze the frequency of distinct infections, meteorological indicators and seasons in children hospitalized for CAP in Salvador, Brazil, nasopharyngeal aspirate and blood were collected from 184 patients aged < 5 y over a 21-month period. Fourteen microbes were investigated and 144 (78%) cases had the aetiology established. Significant differences were found in air temperature between spring and summer (p = 0.02) or winter (p < 0.001), summer and fall (p = 0.007) or winter (p < 0.001), fall and winter (p = 0.002), and on precipitation between spring and fall (p = 0.01). Correlations were found between: overall viral infections and relative humidity (p = 0.006; r = 0.6) or precipitation (p = 0.03; r = 0.5), parainfluenza and precipitation (p = 0.02; r = -0.5), respiratory syncytial virus (RSV) and air temperature (p = 0.048; r = -0.4) or precipitation (p = 0.045; r = 0.4), adenovirus and precipitation (p = 0.02; r = 0.5), pneumococcus and air temperature (p = 0.04; r = -0.4), and Chlamydia trachomatis and relative humidity (p = 0.02; r = -0.5). The frequency of parainfluenza infection was highest during spring (32.1%; p = 0.005) and that of RSV infection was highest in the fall (36.4%; p < 0.001). Correlations at regular strength were found between several microbes and meteorological indicators. Parainfluenza and RSV presented marked seasonal patterns.

Overactivation of glutamate receptors results in neurodegeneration in a variety of brain pathologies, including ischemia, epilepsy, traumatic brain injury and slow-progressing neurodegenerative disorders. In all these pathologies, it is well accepted that the calcium-dependent cysteine proteases calpains are key players in the mechanisms of neuronal cell death. Many research groups have been actively pursuing to establish a link between the deregulation of intracellular Ca(2+) homeostasis associated with excitotoxicity and calpain activity. It is well established that these two events are connected and interact synergistically to promote neurodegeneration, but whether calpain activity depends on or contributes to Ca(2+) deregulation is still under debate.

We report the selection and amplification of the broad-host-range Salmonella phage phi PVP-SE1 in an alternative nonpathogenic host. The lytic spectrum and the phage DNA restriction profile were not modified upon replication in Escherichia coli Bl21, suggesting the possibility of producing this phage in a nonpathogenic host, contributing to the safety and easier approval of a product based on this Salmonella biocontrol agent.

Background: Community-acquired pneumonia (CAP) is a leading cause of childhood death. There are few published reports of radiographic findings among children with severe CAP., Objective: To describe chest X-ray (CXR) findings and assess association between these radiographic findings and pneumococcal isolation in children with severe CAP., Methods: A prospective, multicenter, observational study was conducted in 12 centers in Argentina, Brazil, and the Dominican Republic. Children aged 3-59 months, hospitalized with severe pneumonia, were included. On admission, blood and pleural effusion cultures were performed. Streptococcus pneumoniae was identified according to standard procedures in the respective national reference laboratory. Chest X-rays were taken on admission and read before the culture results were reported., Results: Out of 2,536 enrolled patients, 283 (11.2%) had S. pneumoniae isolated, in 181 cases (7.1%) from blood. The follow radiographic patterns were observed: alveolar infiltrate (75.2%), pleural effusion (15.6%), and interstitial infiltrate (9.2%). Overall, pleural effusion was associated with pneumococcal isolation and pneumococcal bacteremia (P < 0.001). Infiltrates were unilateral (78.7%) or bilateral (21.3%), right-sided (76%) or left-sided (24%), in the lower lobe (53.6%) or the upper lobe (46.4%). Multivariate analysis including patients with affection of only one lobe showed that upper lobe affection and pleural effusion were associated with pneumococcal isolation (OR 1.8, 95% CI, 1.3-2.7; OR 11.0, 95% CI, 4.6-26.8, respectively) and with pneumococcal bacteremia (OR 1.7, 95% CI, 1.2-2.6; OR 3.1, 95% CI, 1.2-8.0, respectively)., Conclusions: Three-quarters of the patients studied had alveolar infiltrates. Upper lobe compromising and pleural effusion were associated with pneumococcal invasive disease., (© 2010 Wiley-Liss, Inc.)

Specific reversed-phase high-performance liquid chromatography conditions are reported for the analysis of recombinant and native human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) preparations. Heterodimeric hLH, hCG and their alpha- and beta-subunits migrated with significantly different retention times (t(R)) in the following order of increasing hydrophobicity: alpha-hCG

Empirical antibiotic use is prescribed in managing children with pneumonia worldwide. We assessed the usefulness of procalcitonin (PCT) and interferon-alpha (IFN-alpha) in differentiating viral from bacterial pneumonia. Among 159 hospitalized children, pneumonia was diagnosed based on clinical complaints plus pulmonary infiltrate. Aetiology was investigated for 9 viruses and 4 atypical and 3 typical bacteria. PCT and IFN-alpha were measured in the serum sample collected on admission. Eight patients had bacteraemic infections, 38 had non-bacteraemic typical infections, and 19 patients had atypical bacterial infections. Viral and unknown aetiology was established in 57 (36%) and 34 (21%) cases, respectively. Three patients with bacterial infection without collected blood culture were excluded. IFN-alpha (IU/ml) was detectable in 20 (13%) cases. The difference among median PCT values of the bacteraemic (4.22; 1.56-7.56), non-bacteraemic typical bacterial (1.47; 0.24-4.07), atypical bacterial (0.18; 0.06-1.03) and only viral (0.65; 0.11-2.22) subgroups was significant (p = 0.02). PCT was > or =2 ng/ml in 52 (33%) cases. The presence of IFN-alpha was associated with PCT <2 ng/ml (90% vs. 64%, p = 0.02). The negative predictive value (95% confidence interval) of PCT > or =2 ng/ml was 95% (89-100%), 89% (78-100%), 93% (85-100%) for differentiation of bacteraemic from viral, atypical bacterial and non-bacteraemic typical bacterial infection, respectively, and 58% (49-68%) for differentiation between bacterial and viral infection. PCT may be useful in identifying bacteraemia among children hospitalized with community-acquired pneumonia. IFN-alpha was uncommonly detected.

Background: Poultry meat is one of the most important sources of human campylobacteriosis, an acute bacterial enteritis which is a major problem worldwide. Campylobacter coli and Campylobacter jejuni are the most common Campylobacter species associated with this disease. These pathogens live in the intestinal tract of most avian species and under commercial conditions they spread rapidly to infect a high proportion of the flock, which makes their treatment and prevention very difficult. Bacteriophages (phages) are naturally occurring predators of bacteria with high specificity and also the capacity to evolve to overcome bacterial resistance. Therefore phage therapy is a promising alternative to antibiotics in animal production. This study tested the efficacy of a phage cocktail composed of three phages for the control of poultry infected with C. coli and C. jejuni. Moreover, it evaluated the effectiveness of two routes of phage administration (by oral gavage and in feed) in order to provide additional information regarding their future use in a poultry unit., Results: The results indicate that experimental colonisation of chicks was successful and that the birds showed no signs of disease even at the highest dose of Campylobacter administered. The phage cocktail was able to reduce the titre of both C. coli and C. jejuni in faeces by approximately 2 log10 cfu/g when administered by oral gavage and in feed. This reduction persisted throughout the experimental period and neither pathogen regained their former numbers. The reduction in Campylobacter titre was achieved earlier (2 days post-phage administration) when the phage cocktail was incorporated in the birds' feed. Campylobacter strains resistant to phage infection were recovered from phage-treated chickens at a frequency of 13%. These resistant phenotypes did not exhibit a reduced ability to colonize the chicken guts and did not revert to sensitive types., Conclusions: Our findings provide further evidence of the efficacy of phage therapy for the control of Campylobacter in poultry. The broad host range of the novel phage cocktail enabled it to target both C. jejuni and C. coli strains. Moreover the reduction of Campylobacter by approximately 2 log10cfu/g, as occurred in our study, could lead to a 30-fold reduction in the incidence of campylobacteriosis associated with consumption of chicken meals (according to mathematical models). To our knowledge this is the first report of phage being administered in feed to Campylobacter-infected chicks and our results show that it lead to an earlier and more sustainable reduction of Campylobacter than administration by oral gavage. Therefore the present study is of extreme importance as it has shown that administering phages to poultry via the food could be successful on a commercial scale.

Objectives: To evaluate patients treated for traumatic injuries and to identify adherence to guidelines recommendations of treatment and association with death. The recommendations adopted were defined by the committee on trauma of the American College of Surgeons in advanced trauma life support., Methods: Retrospective cohort study conducted at a teaching hospital. The study population was victims of trauma > 12 years of age with injury severity scores > 16 who were treated between January 1997 and December 2001. Data collection was divided into three phases: pre-hospital, in-hospital, and post-mortem. The data collected were analyzed using EPI INFO., Results: We analyzed 207 patients, 147 blunt trauma victims (71%) and 60 (29%) penetrating trauma victims. Trauma victims had a 40.1% mortality rate. We identified 221 non adherence events that occurred in 137 patients. We found a mean of 1.61 non adherence per patient, and it occurred less frequently in survivors (1.4) than in non-survivors (1.9; p=0.033). According to the trauma score and injury severity score methodology, 54.2% of deaths were considered potentially preventable. Non adherence occurred 1.77 times more frequently in those considered potentially preventable deaths compared to other non-survivors (95% CI: 1.12-2.77; p=0.012), and 92.9% of the multiple non adherence occurred in the first group (p=0.029)., Conclusions: Non adherence occurred more frequently in patients with potentially preventable deaths. Non adherence to guidelines recommendations can be considered a contributing factor to death in trauma victims and can lead to an increase in the number of potentially preventable deaths.

Objective: The aim of this study was to assess and compare the effects of laser phototherapy (LPT) on cutaneous flaps on diabetic rats., Background: Diabetes mellitus is characterized by high blood glucose levels. Its main complications are delayed wound healing, an impaired blood supply, and a decrease in collagen production. Cutaneous flaps are routinely used in several surgical procedures, and most failures are related to poor blood supply. LPT has been studied using several healing models., Animals and Methods: Twelve Wistar rats were randomized into three groups: group 1 (G1; diabetic animals without treatment), group 2 (G2; diabetic animals irradiated with lambda680 nm), and group 3 (G3; diabetic animals irradiated with lambda790 nm). Diabetes was induced with streptozotocin. A 2- x 8-cm cutaneous flap was raised on the dorsum of each animal, and a plastic sheet was introduced between the flap and the bed to cause poor blood supply. Nonirradiated animals acted as controls. The dose per session was 40 J/cm(2). Laser light was applied transcutaneously and fractioned on 16 contact points at the wound margins (16 x 2.5 J/cm(2)). Animal death occurred on day 8 after surgery. Specimens were taken, processed, cut, stained with eosin (HE) and sirius red, and underwent histological analysis., Results: It is shown that accute inflammation was mostly discrete for G3. Chronic inflammation was more evident for G2. Fibroblast number was higher for G3. Angiogenesis was more evident for G3. Necrosis was more evident for G2. Statistical analysis among all groups showed significant differences (p = 0.04) on the level of acute inflammation between G1 and G3, tissue necrosis between G1 and G2 (p = 0.03), chronic inflammation between (p = 0.04), fibroblastic proliferation between G2 and G3 (p = 0.05), and neovascularization between G2 and G3 (p = 0.04)., Conclusion: LPT was effective in increasing angiogenesis as seen on irradiated subjects and was more pronounced when IR laser light was used.

Photodynamic therapy has been used to inactivate microorganisms through the use of targeted photosensitizers. Although the photoinactivation of microorganisms has already been studied under different conditions, a systematic evaluation of irradiation characteristics is still limited. The goal of this study was to test how the light dose, fluence rate and irradiation source affect the viral photoinactivation of a T4-like sewage bacteriophage. The experiments were carried out using white PAR light delivered by fluorescent PAR lamps (40 W m(-2)), sun light (600 W m(-2)) and an halogen lamp (40-1690 W m(-2)). Phage suspensions and two cationic photosensitizers (Tetra-Py(+)-Me, Tri-Py(+)-Me-PF) at concentrations of 0.5, 1.0 and 5.0 microM were used. The results showed that the efficacy of the bacteriophage photoinactivation is correlated not only with the sensitizer and its concentration but also with the light source, energy dose and fluence rate applied. Both photosensitizers at 5.0 microM were able to inactivate the T4-like phage to the limit of detection for each light source and fluence rate. However, depending of the light parameters, different irradiation times are required. The efficiency of photoinactivation is dependent on the spectral emission distribution of the light sources used. Considering the same light source and a fixed light dose applied at different fluence rates, phage inactivation was significantly higher when low fluence rates were used. In this way, the light source, fluence rate and total light dose play an important role in the effectiveness of the antimicrobial photodynamic therapy and should always be considered when establishing an optimal antimicrobial protocol.

Nitric oxide (NO) was described to inhibit the proliferation of neural stem cells. Some evidence suggests that NO, under certain conditions, can also promote cell proliferation, although the mechanisms responsible for a potential proliferative effect of NO in neural stem cells have remained unaddressed. In this work, we investigated and characterized the proliferative effect of NO in cell cultures obtained from the mouse subventricular zone. We found that the NO donor NOC-18 (10 microM) increased cell proliferation, whereas higher concentrations (100 microM) inhibited cell proliferation. Increased cell proliferation was detected rapidly following exposure to NO and was prevented by blocking the mitogen-activated kinase (MAPK) pathway, independently of the epidermal growth factor (EGF) receptor. Downstream of the EGF receptor, NO activated p21Ras and the MAPK pathway, resulting in a decrease in the nuclear presence of the cyclin-dependent kinase inhibitor 1, p27(KIP1), allowing for cell cycle progression. Furthermore, in a mouse model that shows increased proliferation of neural stem cells in the hippocampus following seizure injury, we observed that the absence of inducible nitric oxide synthase (iNOS(-/-) mice) prevented the increase in cell proliferation observed following seizures in wild-type mice, showing that NO from iNOS origin is important for increased cell proliferation following a brain insult. Overall, we show that NO is able to stimulate the proliferation of neural stem cells bypassing the EGF receptor and promoting cell division. Moreover, under pathophysiological conditions in vivo, NO from iNOS origin also promotes proliferation in the hippocampus.

Context: Osteoporosis is a skeletal disorder characterized by low bone mass and microarchitectural deterioration of bone tissue. It may affect the craniomaxillofacial complex and result in less bone-implant contact., Aims: The objective of this study was to measure the endosseous integration of titanium implants using the removal torque test in low-mineral density experimental animal models., Materials and Methods: Thirty female New Zealand white rabbits were divided into two groups: control (n=14), sham-operated; and test (n=16), ovariectomized. All animals received 2.2 mm Χ 4.0 mm titanium implants in the right tibia, 120 days after ovariectomy or sham surgery. Animals were killed at 7 and 14 weeks of postimplant insertion, when implants were removed by reverse torque rotation using a digital torque meter. Statistical analysis used was Student's t-test., Results and Conclusion: At the 7-week period, the mean torque values for the control and test groups were 11.6 Ncm and 10.4 Ncm, respectively, with no statistical significance. Implant removal torque analysis showed statistically lower values (P < 0.05) for the test group at the 14-week period (control=16.7 Ncm and test=9.7 Ncm). The results of this study indicate that hormonal privacy compromise implants biomechanical performance.

Epizootiological study of Anaplasma marginale in regions that contain various reservoir hosts, co-existence of rickettsia pathogens, and common vectors is a complicated task. To achieve diagnosis of this rickettsia in cattle and campeiro deer of Brazilian Pantanal, a comparison was made between a real time polymerase chain reaction (RT-PCR) with intercalating Sybr Green fluorochrome and primers based on msp5 gene of A. marginale; a conventional PCR (C-PCR); and parasitological examination using thin blood smear stained with Giemsa-MayGrunwald. Both PCRs showed good performance in the diagnosis of A. marginale in cattle, and were superior to the parasitological exam. The RT-PCR detected seven positive campeiro deer (16.3%). This rate was significantly higher compared to C-PCR, which identified one animal as positive (2.3%), and also compared to parasitological diagnosis, which did not find any positive animals. The dissociation temperature average of positive reactions in cattle (81.72 °C ± 0.20) was identical to dissociation temperature found in the cervids (81.72 °C ± 0.12), suggesting that both animal species were infected with A. marginale. We concluded that RT-PCR can be used for A. marginale diagnosis and in epizootiological studies of cattle and cervids; in spite of the small number of campeiro deer samples, the results indicated that this wildlife species has importance in the Anaplasma epizootiology in the Brazilian Pantanal.

Objective: To estimate the clinical failure and adverse events in children with nonsevere pneumonia receiving amoxicillin, identifying risk factors., Research Design/methods: 192 patients aged 2 - 59 months were prospectively followed up. Pneumonia diagnosis was based on respiratory complaints and radiographic pulmonary infiltrate or pleural effusion. Amoxicillin (50 mg/kg/day) was given. Demographic data and clinical findings on admission, daily evolution up to the 5th day of treatment and 2 - 4 weeks after enrollment were collected., Main Outcome Measures: Clinical failure included persistence of fever, difficulty breathing or tachypnea beyond the first 48 h of treatment or of cough beyond the first 96 h of treatment or sign of severe/very severe disease up to the 5th day of treatment., Results: Amoxicillin failed in 6 (3.1%) cases. By excluding one child diagnosed with cystic fibrosis after continued follow-up, the final clinical failure rate was 2.6%. The total adverse effect frequency was 14 (7.3%), but amoxicillin was discontinued only in 1 (0.5%) case. No relapse was identified at the 2 - 4-week interval evaluation. By multivariate analysis, age (OR = 1.1; 95% CI 1.01 - 1.19) was an independent risk factor for clinical failure which occurred in older children (47 +/- 9 vs 31 +/- 16 months; p = 0.01)., Conclusions: Clinical failures were few, especially among those aged < 2 years. Amoxicillin discontinuation due to adverse reaction was rare.

A qualitative research was realized to verify the occurrence and motivations for self-medication and the quality of the information rendered by pharmacies. We conducted 3 focus groups with 25 patients of a reference public health service for STD treatment in Brasília, Brazil. We used the critical discourse analysis to interpret the data obtained during interviews. The analysis revealed that self-medication was a common practice among participants, motivated mainly by discontentment with the long waiting period and the quality of the public health services. Other motivations included: previous experience with medications, advice from friends and family members and the search for an anonymous service provided at pharmacies due to embarrassment and inadequate care environment at health care services. Care provided at pharmacies was also considered unsatisfactory. Viewed essentially as a commercial establishment, pharmacies have become, yet, an easy accessible alternative. Most of participants perceived the pharmacists as ordinary employees seeking only to increase profits. The results suggest a need to improve access to health care services, as well as to render it more humane. Community pharmacies should to be turned into health establishments to participate in health education and orientation for rational drugs use.

Temporomandibular disorders (TMDs) are common painful multifactorial conditions affecting the temporomandibular joint (TMJ) and whose treatment depends on the type and symptoms. Initially, it requires pain control, and, for this, drugs, biting plates, occlusal adjustment, physiotherapy or their association are used. Lately, laser phototherapy (LPT) has been used in the treatment of pain of several origins, including TMDs. This study reports the treatment of a selected group of 74 patients treated at the Laser Center of the Federal University of Bahia between 2003 and 2008. Following standard anamneses, clinical and imaging examination and with the diagnosis of any type of TMD, the patients were prepared for LPT. No other intervention was carried out during the treatment. Treatment consisted of three sessions a week for 6 weeks. Prior to irradiation, the patients were asked to score their pain using a visual analog scale (VAS). Lasers of wavelength (lambda)780 nm, lambda790 nm or lambda830 nm and/or lambda660 nm were used at each session (30/40 mW; spot (varphi) approximately 3 mm; mean dose per session 14.2 +/- 6.8 J/cm(2); mean treatment dose of 170 +/- 79.8 J/cm(2)). Of the patients, 80% were female ( approximately 46 years old). At the end of the 12 sessions the patients were again examined, and they scored their pain using the VAS. The results were statistically analyzed and showed that 64% of the patients were asymptomatic or had improved after treatment and that the association of both wavelengths was statistically significant (P = 0.02) in the asymptomatic group. It was concluded that the association of red and infrared (IR) laser light was effective in pain reduction on TMJ disorders of several origins.

Importance of the Field: Community-acquired pneumonia (CAP) is a leading cause of morbidity and mortality among children under 5 years. Although viral infections have been frequently recognized, death has been attributable to bacterial infections (especially due to Streptococcus pneumoniae)., Areas Covered in This Review: In the past twenty years, childhood mortality due to CAP has been declining because of accessible health care and prompt antibiotic use. The basic options of antibiotics to treat children with CAP are presented., What the Reader Will Gain: In daily practice, antibiotic prescription to treat childhood CAP is based on age and severity assessment. For those children aged less than 2 months, hospitalization and parenteral administration of ampicillin plus aminoglycoside or ampicillin plus third-generation cephalosporin is mandatory. For those patients aged 2 months and older with non-severe or severe CAP, the first option is amoxicillin or aqueous penicillin G, respectively; for very severe presentation, oxacillin and third-generation cephalosporin are recommended. If atypical bacteria infection is suspected, macrolide must be given in any age or severity group., Take Home Message: Few antibiotics are included in the guidelines to treat childhood CAP. The choice of which antibiotic should be given is based on clinical evaluation.

Aim: The aim of the present study was to assess histologically the effect of Laser Photobiomodulation (LPBM) on skin wounds treated with Mitomycin C (MMC)., Background Data: Wound healing occurs because of a competitive mechanism between the synthesis and lyses of collagen. Therefore, any factor that increases the lyses or reduces the synthesis of collagen may result in changes in the healing process. MMC is an antineoplastic drug that inhibits fibroblast proliferation, collagen synthesis, and neoangiogenesis. LPBM has been shown to stimulate wound healing, increasing the production of collagen, fibroblastic proliferation, and angiogenesis., Materials and Methods: Forty-eight Wistar rats were randomly distributed into 4 main groups (n = 12): G1--control (G1a--7 d and G1b--14 d); G2--MMC (G2a--7 d and G2b--14 d); G3--MMC + lambda660 nm laser (G3a--7 d and G3b--14 d); and G4--MMC + lambda790 nm laser (G4a--7 d and G4b--14 d). Under general anesthesia, one excisional wound was created on the dorsum of each animal. Two ml of MMC solution was applied to the wound 4 h after surgery for 5 min. LPBM was performed on groups G3 (lambda690 nm; 20 J/cm(2); 30 mW; Phi = 2 mm) and G4 (lambda790 nm; 20 J/cm(2); 40 mW; Phi = 2 mm), starting immediately after the application of the MMC and repeated every other day during the experimental period. Laser light was applied transcutaneously at 4 equidistant points on the wound margin (4 x 5 J/cm(2), 20 J/cm(2)/session). The specimens were routinely cut and processed to wax. The slides were stained with HE and Sirius red. Computerized hystomorphometry was performed., Results: LPBM resulted in reduced inflammation and an increase in both fibroblast proliferation and collagen deposition., Conclusion: The use of LPBM improves wound healing in subjects treated with MMC.

Gene duplications alter the genetic constitution of organisms and can be a driving force of molecular evolution in humans and the great apes. In this context, the study of genomic disorders has uncovered the essential role played by the genomic architecture, especially low copy repeats (LCRs) or segmental duplications (SDs). In fact, regardless of the mechanism, LCRs can mediate or stimulate rearrangements, inciting genomic instability and generating dynamic and unstable regions prone to rapid molecular evolution. In humans, copy-number variation (CNV) has been implicated in common traits such as neuropathy, hypertension, color blindness, infertility, and behavioral traits including autism and schizophrenia, as well as disease susceptibility to HIV, lupus nephritis, and psoriasis among many other clinical phenotypes. The same mechanisms implicated in the origin of genomic disorders may also play a role in the emergence of segmental duplications and the evolution of new genes by means of genomic and gene duplication and triplication, exon shuffling, exon accretion, and fusion/fission events.

Antimicrobial photodynamic therapy (aPDT) has emerged in the clinical field as a potential alternative to antibiotics to treat microbial infections. No cases of microbial viability recovery or any resistance mechanisms against it are yet known. 5,10,15-tris(1-Methylpyridinium-4-yl)-20-(pentafluorophenyl)-porphyrin triiodide (Tri-Py(+)-Me-PF) was used as photosensitizer. Vibrio fischeri and recombinant Escherichia coli were the studied bacteria. To determine the bacterial recovery after treatment, Tri-Py(+)-Me-PF (5.0 microM) was added to bacterial suspensions and the samples were irradiated with white light (40 W m(-2)) for 270 minutes. Then, the samples were protected from light, aliquots collected at different intervals and the bioluminescence measured. To assess the development of resistance after treatment, bacterial suspensions were exposed to white light (25 minutes), in presence of 5.0 microM of Tri-Py(+)-Me-PF (99.99% of inactivation) and plated. After the first irradiation period, surviving colonies were collected from the plate and resuspended in PBS. Then, an identical protocol was used and repeated ten times for each bacterium. The results suggest that aPDT using Tri-Py(+)-Me-PF represents a promising approach to efficiently destroy bacteria since after a single treatment these microorganisms do not recover their viability and after ten generations of partially photosensitized cells neither of the bacteria develop resistance to the photodynamic process.

An inducible system has been established in Nicotiana tabacum plants allowing controlled expression of Sar1-GTP and thus the investigation of protein dynamics after inhibition of endoplasmic reticulum (ER) to Golgi transport. Complete Golgi disassembly and redistribution of Golgi markers into the ER was observed within 18-24h after induction. At the ultrastructural level Sar1-GTP expression led to a decrease in Golgi stack size followed by Golgi fragmentation and accumulation of vesicle remnants. Induction of Sar1-GTP resulted in redistribution of the green fluorescent protein (GFP)-tagged Arabidopsis golgins AtCASP and GC1 (golgin candidate 1, an Arabidopsis golgin 84 isoform) into the ER or cytoplasm, respectively. Additionally, both fusion proteins were observed in punctate structures, which co-located with a yellow fluorescent protein (YFP)-tagged version of Sar1-GTP. The Sar1-GTP-inducible system is compared with constitutive Sar1-GTP expression and brefeldin A treatment, and its potential for the study of the composition of ER exit sites and early cis-Golgi structures is discussed.

Objective: There have been no objective assessments to determine whether boys with MECP2 duplication have autism or whether female carriers manifest phenotypes. This study characterizes the clinical and neuropsychiatric phenotypes of affected boys and carrier females., Methods: Eight families (9 males and 9 females) with MECP2 duplication participated. A detailed history, physical examination, electroencephalogram, developmental evaluation, Autism Diagnostic Observation Schedule, and Autism Diagnostic Interview-Revised were performed for each boy. Carrier females completed the Symptom Checklist-90-R, Wechsler Abbreviated Scale of Intelligence, Broad Autism Phenotype Questionnaire, and detailed medical and mental health histories. Size and gene content of each duplication were determined by array comparative genome hybridization. X-chromosome inactivation patterns were analyzed using leukocyte DNA. MECP2 and IRAK1 RNA levels were quantified from lymphoblast cell lines, and western blots were performed to assess MeCP2 protein levels., Results: All of the boys demonstrated mental retardation and autism. Poor expressive language, gaze avoidance, repetitive behaviors, anxiety, and atypical socialization were prevalent. Female carriers had psychiatric symptoms, including generalized anxiety, depression, and compulsions that preceded the birth of their children. The majority exhibited features of the broad autism phenotype and had higher nonverbal compared to verbal reasoning skills., Interpretation: Autism is a defining feature of the MECP2 duplication syndrome in boys. Females manifest phenotypes despite 100% skewing of X-inactivation and normal MECP2 RNA levels in peripheral blood. Analysis of the duplication size, MECP2 and IRAK1 RNA levels, and MeCP2 protein levels revealed that most of the traits in affected boys are likely due to the genomic region spanning of MECP2 and IRAK1. The phenotypes observed in carrier females may be secondary to tissue-specific dosage alterations and require further study. Ann Neurol 2009;66:771-782.

Aim: We evaluated histologically the differences in the healing of cutaneous wounds on nourished or undernourished rats following laser phototherapy (LPT) (20 or 40 J/cm2)., Background: Wound healing occurs due to a competitive mechanism between the synthesis and lysis of collagen. Any factor that increases the lysis or reduces the synthesis of collagen may change the healing process. Nutritional deficiencies have a great effect on wound healing., Methods: Fifty nourished or undernourished Wistar rats had one standardized wound created on the dorsum and were divided into 10 subgroups: Control (standard diet); Control [Northeastern Brazilian basic diet (DBR)]; Standard diet+LPT (lambda635 nm; spatial average energy fluence (SAEF) 20 J/cm2); Standard diet+LPT (lambda635 nm; SAEF 40 J/cm2); Standard diet+LPT (lambda780 nm; SAEF 20 J/cm2); Standard diet+LPT (lambda780 nm; SAEF 40 J/cm2); DBR+LPT (lambda635 nm; SAEF 20 J/cm2); DBR+LPT (lambda635 nm; SAEF 40 J/cm2); DBR+LPT (lambda780 nm; SAEF 20 J/cm2); DBR+LPT (lambda780 nm; SAEF 40 J/cm2). The first application of the treatment was carried out immediately and repeated daily for seven days. The specimens were routinely processed to wax, cut, stained with H&E and Sirius Red stains, and analyzed using light microscopy. The analyses included re-epithelization, inflammatory infiltrate, and fibroblastic proliferation. Sirius Red stained slides were used to perform descriptive analyses of the collagen fibers., Results: The results showed that LPT with lambda635 wavelength was more effective for the treatment of undernourished subjects, treated with either LPT with lambda730 nm with SAEF of 20 J/cm2 or lambda780 nm with SAEF of 40 J/cm2., Conclusion: Nutritional status influenced the progression of the healing process as well as the quality of the healed tissue. In addition, the use of both wavelengths resulted in a positive biomodulatory effect on both nourished and undernourished subjects.

We performed an inventory of soybean NAC transcription factors, in which 101 NAC domain-containing proteins were annotated into 15 different subgroups, showing a clear relationship between structure and function. The six previously described GmNAC proteins (GmNAC1 to GmNAC6) were located in the nucleus and a transactivation assay in yeast confirmed that GmNAC2, GmNAC3, GmNAC4 and GmNAC5 function as transactivators. We also analyzed the expression of the six NAC genes in response to a variety of stress conditions. GmNAC2, GmNAC3 and GmNAC4 were strongly induced by osmotic stress. GmNAC3 and GmNAC4 were also induced by ABA, JA and salinity but differed in their response to cold. Consistent with an involvement in cell death programs, the transient expression of GmNAC1, GmNAC5 and GmNAC6 in tobacco leaves resulted in cell death and enhanced expression of senescence markers. Our results indicate that the described soybean NACs are functionally non-redundant transcription factors involved in response to abiotic stresses and in cell death events in soybean.

Objective: This study aimed to analyze outcome, clinical and epidemiological characteristics and severity factors in adult patients admitted with a diagnosis of infection by virus A (H1N1) to public and private intensive care units, in Paraná, Brazil., Methods: Cohort study of medical charts of patients older than 12 years admitted to 11 intensive care units in 6 cities in the state of Parana, Brazil, during a period of 45 days, with diagnosis of swine influenza. The diagnosis of infection with A (H1N1) was made by real time polymerase chain reaction (RT-PCR) of nasopharyngeal secretion, or strong clinical suspicion when other causes had been ruled out (even with negative RT-PCR). Descriptive statistics were performed, analysis by the Chi square test was used to compare percentages and the Student's t test for continuous variables with univariate analysis, assuming a significance level of p <0.05., Results: There were 63 adult patients admitted with a diagnosis of H1N1, 37 (58.7%) being RT-PCR positive. Most patients were young adults (65% under 40 years of age) with no gender predominance and high incidence of obesity (27.0% with Body Mass Index > 30). Mean of the Acute Physiologic Chronic Health Evaluation II (APACHE II) score was 15.0 + 8.1. Mortality in the intensive care unit was 39.7%. The main factors associated with mortality were: positive RT-PCR, low levels of initial PaO2/FiO2, high initial levels of urea and lactate dehydrogenase, required level of positive end expiratory pressure, need for the prone position and vasopressors., Conclusions: Adult patients with A (H1N1) virus infection admitted to intensive care units had a high risk of death, particularly due to respiratory impairment. Positive RT-PCR, urea and lactic dehydrogenase, low initial PaO2/FiO2 and high levels of PEEP were correlated with higher mortality.

Peripheral tethering factors bind to small GTPases in order to obtain their correct location within the Golgi apparatus. Using fluorescence resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) we visualized interactions between Arabidopsis homologues of tethering factors and small GTPases at the Golgi stacks in planta. Co-expression of the coiled-coil proteins AtGRIP and golgin candidate 5 (GC5) [TATA element modulatory factor (TMF)] and the putative post-Golgi tethering factor AtVPS52 fused to green fluorescent protein (GFP) with mRFP (monomeric red fluorescent protein) fusions to the small GTPases AtRab-H1(b), AtRab-H1(c) and AtARL1 resulted in reduced GFP lifetimes compared to the control proteins. Interestingly, we observed differences in GFP quenching between the different protein combinations as well as selective quenching of GFP-AtVPS52-labelled structures. The data presented here indicate that the FRET-FLIM technique should prove invaluable in assessing protein interactions in living plant cells at the organelle level.

Background: The Double-Layer Agar (DLA) technique is extensively used in phage research to enumerate and identify phages and to isolate mutants and new phages. Many phages form large and well-defined plaques that are easily observed so that they can be enumerated when plated by the DLA technique. However, some give rise to small and turbid plaques that are very difficult to detect and count. To overcome these problems, some authors have suggested the use of dyes to improve the contrast between the plaques and the turbid host lawns. It has been reported that some antibiotics stimulate bacteria to produce phages, resulting in an increase in final titer. Thus, antibiotics might contribute to increasing plaque size in solid media., Results: Antibiotics with different mechanisms of action were tested for their ability to enhance plaque morphology without suppressing phage development. Some antibiotics increased the phage plaque surface by up to 50-fold., Conclusion: This work presents a modification of the DLA technique that can be used routinely in the laboratory, leading to a more accurate enumeration of phages that would be difficult or even impossible otherwise.