Occurrence of non-O157 Shiga toxin-producing Escherichia coli in dogs with diarrhea

Shiga toxigenic Escherichia coli (STEC) and Attaching and effacing E. coli (AEEC) have been associated with diarrhea illness in dogs. From January to December 2006, 92 E. coli isolates from 25 diarrheic dogs were analyzed, by screening for the presence of Shiga toxin-producing (stx 1 and stx 2) and intimin (eae) genes. Twelve isolates were detected by PCR to harbor the Shiga toxin genes (7 the stx 1 (7.6%); 5 the stx 2 (5.4%); and none both of them). Nine (9.8%) of the E. coli isolates studied were eae positive non Shiga toxin-producing. Thirteen (62.0%) isolates, carrying stx or eae gene, also showed a hemolysin production. The strains with virulence genes were also examined for resistance to 12 antimicrobial agents. Resistances to cephalothin (85.7%), streptomycin (81.0%), amoxicillin (71.4%) and gentamicin (71.4%) were predominantly observed.

Characterization of aqueous silver nitrate solutions for leakage tests

OBJECTIVE: To determine the pH over a period of 168 h and the ionic silver content in various concentrations and post-preparation times of aqueous silver nitrate solutions. Also, the possible effects of these factors on microleakage test in adhesive/resin restorations in primary and permanent teeth were evaluated. MATERIAL AND METHODS: A digital pHmeter was used for measuring the pH of the solutions prepared with three types of water (purified, deionized or distilled) and three brands of silver nitrate salt (Merck, Synth or Cennabras) at 0, 1, 2, 24, 48, 72, 96 and 168 h after preparation, and storage in transparent or dark bottles. Ionic silver was assayed according to the post-preparation times (2, 24, 48, 72, 96, 168 h) and concentrations (1, 5, 25, 50%) of solutions by atomic emission spectrometry. For each sample of each condition, three readings were obtained for calculating the mean value. Class V cavities were prepared with enamel margins on primary and permanent teeth and restored with the adhesive systems OptiBond FL or OptiBond SOLO Plus SE and the composite resin Filtek Z-250. After nail polish coverage, the permanent teeth were immersed in 25% or 50% AgNO3 solution and the primary teeth in 5% or 50% AgNO3 solutions for microleakage evaluation. ANOVA and the Tukey's test were used for data analyses (α=5%). RESULTS: The mean pH of the solutions ranged from neutral to alkaline (7.9±2.2 to 11.8±0.9). Mean ionic silver content differed depending on the concentration of the solution (4.75±0.5 to 293±15.3 ppm). In the microleakage test, significant difference was only observed for the adhesive system factor (p=0.000). CONCLUSIONS: Under the tested experimental conditions and based on the obtained results, it may be concluded that the aqueous AgNO3 solutions: have neutral/alkaline pH and service life of up to 168 h; the level of ionic silver is proportional to the concentration of the solution; even at 5% concentration, the solutions were capable of indicating loss of marginal seal in the composite restorations; the 3-step conventional adhesive system had better performance regarding microleakage in enamel on primary and permanent teeth.

Valores de referência e influência da idade no eritrograma de bubalinos da raça Murrah

O eritrograma é uma avaliação que representa um auxílio ao clínico no diagnóstico de afecções que acometem os animais domésticos. No entanto, estudos hematológicos em bubalinos são escassos, havendo poucas informações na literatura a respeito de valores de referência dos constituintes sanguíneos desta espécie. Assim, este trabalho teve como objetivo a análise do eritrograma de búfalos de diferentes faixas etárias da raça Murrah, machos e fêmeas. Os animais em estudo foram distribuídos em quatro grupos experimentais, de acordo com as idades: Grupo 1: animais com idade entre o nascimento e 3 meses (n=15); Grupo 2: animais com idade entre 4 e 6 meses (n=50); Grupo 3: animais com idade entre 7 e 12 meses (n=50); e Grupo 4: animais com idade superior a 12 meses (n=50). Como resultados das análises foram encontrados os seguintes valores médios, no Grupo 1: 7,9x106 hemácias/mL de sangue (He); 13,0 g/dL de hemoglobina (Hb); hematócrito (Ht) de 38,9%; Volume Corpuscular Médio (VCM) de 49,0 fl; Concentração Hemoglobínica Corpuscular Média (CHCM) de 33,6 %, e Hemoglobina Corpuscular Média (HCM) de 16,4 pg; no Grupo 2: He: 7,1 x10(6)/mL; Hb: 12,5 g/dL; Ht: 36,8%; VCM: 52,4 fl; CHCM: 33,9%, e HCM: 17,8 pg; no Grupo 3: He: 7,9 x10(6)/mL; Hb: 12,0 g/dL; Ht: 33,8%; VCM: 43,1 fl; CHCM: 35,4%, e HCM: 15,34 pg; e no Grupo 4: He: 6,7 x10(6)/mL; Hb: 11,7 g/dL; Ht: 34,4%; VCM: 53,4 fl; CHCM: 34,4%. e HCM: 17,4 pg. A análise estatística dos resultados encontrados para as diferentes faixas etárias permitiu concluir que ocorreram variações determinadas pela evolução da idade, caracterizadas por diminuição do número médio de hemácias (He), da concentração de hemoglobina (Hb) e do valor do hematócrito (Ht). Os índices hematimétricos apresentaram variações significativas no G3.

Balance control in hemiparetic stroke patients: Main tools for evaluation

Balance problems in hemiparetic patients after stroke can be caused by different impairments in the physiological systems involved in Postural control, including sensory afferents, movement strategies, biomechanical constraints, cognitive processing, and perception of verticality. Balance impairments and disabilities must be appropriately addressed. This article reviews the most common balance abnormalities in hemiparetic patients with stroke and the main tools used to diagnose them.

Leucine supplementation improves adiponectin and total cholesterol concentrations despite the lack of changes in adiposity or glucose homeostasis in rats previously exposed to a high-fat diet

Background: Studies suggest that leucine supplementation (LS) has a therapeutic potential to prevent obesity and to promote glucose homeostasis. Furthermore, regular physical exercise is a widely accepted strategy for body weight maintenance and also for the prevention of obesity. The aim of this study was to determine the effect of chronic LS alone or combined with endurance training (ET) as potential approaches for reversing the insulin resistance and obesity induced by a high-fat diet (HFD) in rats. Methods: Forty-seven rats were randomly divided into two groups. Animals were fed a control diet-low fat (n = 10) or HFD (n = 37). After 15 weeks on HFD, all rats received the control diet-low fat and were randomly divided according to treatment: reference (REF), LS, ET, and LS+ET (n = 7-8 rats per group). After 6 weeks of treatment, the animals were sacrificed and body composition, fat cell volume, and serum concentrations of total cholesterol, HDL-cholesterol, triacylglycerol, glucose, adiponectin, leptin and tumor necrosis factor-alpha (TNF-alpha) were analyzed. Results: At the end of the sixth week of treatment, there was no significant difference in body weight between the REF, LS, ET and LS+ET groups. However, ET increased lean body mass in rats (P = 0.019). In addition, ET was more effective than LS in reducing adiposity (P = 0.019), serum insulin (P = 0.022) and TNF-alpha (P = 0.044). Conversely, LS increased serum adiponectin (P = 0.021) levels and reduced serum total cholesterol concentration (P = 0.042). Conclusions: The results showed that LS had no beneficial effects on insulin sensitivity or adiposity in previously obese rats. On the other hand, LS was effective in increasing adiponectin levels and in reducing total cholesterol concentration.

Effect of eucalyptus (Eucalyptus citriodora) fresh or residue leaves on methane emission in vitro

Rumen fermentation and methane emission for eucalyptus (Eucalyptus citriodora) fresh leaves (FL) or residue leaves (RL), after essential oil extraction from eucalyptus leaves in comparison with alfalfa (Medicago sativa) hay, were investigated in vitro. Eucalyptus FL and RL were obtained from the Distillery Trees Barras Company, Torrinha City, Sao Paulo, Brazil. The semi-automatic system of gas production was used to measure gas production, methane emission and rumen fermentation after 24 h incubation in vitro. The results showed that the crude protein (CP) contents were 76.4, 78.1 and 181.9 g kg(-1) DM for eucalyptus FL, RL and alfalfa hay, respectively. The neutral-detergent fibre (NDF) and acid-detergent fibre (ADF) were significantly lower in eucalyptus FL and RL than alfalfa hay. The Eucalyptus fresh and residue leaves were rich in total phenols (TP) and total tannins (TT) but had negligible content of condensed tannins (CT). There was significant reduction in cumulative gas production about 54 and 51% with eucalyptus FL and RL, respectively, compared with alfalfa hay. The methane emission (mL/g DM) was reduced (P<0.05) by 53 and 57% with eucalyptus FL and RL, respectively, but the reduction was 21 and 16% when expressed on truly digested organic matter basis. There were a decline (P<0.05) in true dry and organic matter degradation in vitro in eucalyptus FL and RL compared with alfalfa hay substrate. The partitioning factor values were higher (P<0.05) in eucalyptus FL and RL than alfalfa hay. There was no significant difference observed between eucalyptus FL, RL and alfalfa hay in protozoa count. It is concluded that the eucalyptus leaves have potential effect to mitigate CH4 production in vitro, which may be attributed to a decrease in fermentable substrate rather than to a direct effect on methanogenesis.

Route to chaos in a third-order phase-locked loop network

Phase-locked loops (PLLs) are widely used in applications related to control systems and telecommunication networks. Here we show that a single-chain master-slave network of third-order PLLs can exhibit stationary, periodic and chaotic behaviors, when the value of a single parameter is varied. Hopf, period-doubling and saddle-saddle bifurcations are found. Chaos appears in dissipative and non-dissipative conditions. Thus, chaotic behaviors with distinct dynamical features can be generated. A way of encoding binary messages using such a chaos-based communication system is suggested. (C) 2009 Elsevier B.V. All rights reserved.

Comparison of Humanized IgG and FvFc Anti-CD3 Monoclonal Antibodies Expressed in CHO Cells

Two humanized monoclonal antibody constructs bearing the same variable regions of an anti-CD3 monoclonal antibody, whole IgG and FvFc, were expressed in CHO cells. Random and site-specific integration were used resulting in similar expression levels. The transfectants were selected with appropriate selection agent, and the surviving cells were plated in semi-solid medium for capture with FITC-conjugated anti-human IG antibody and picked with the robotic ClonePix FL. Conditioned media from selected clones were purified by affinity chromatography and characterized by SDS-PAGE, Western-blot, SEC-HPLC, and isoelectric focusing. Binding to the target present in healthy human mononuclear cells was assessed by flow cytometry, as well as by competition between the two constructs and the original murine monoclonal antibody. The humanized constructs were not able to dislodge the murine antibody while the murine anti-CD3 antibody could dislodge around 20% of the FvFc or IgG humanized versions. Further in vitro and in vivo pre-clinical analyses will be carried out to verify the ability of the humanized versions to demonstrate the immunoregulatory profile required for a humanized anti-CD3 monoclonal antibody.

Differences Between ""Classical"" Risk Factors for Infections Caused by Methicillin-Resistant Staphylococcus aureus (MRSA) and Risk Factors for Nosocomial Bloodstream Infections Caused by Multiple Clones of the Staphylococcal Cassette Chromosome mec Type IV MRSA Strain

OBJECTIVE. To identify risk factors associated with nosocomial bloodstream infections caused by multiple clones of the staphylococcal cassette chromosome mec (SCCmec) type IV strain of methicillin-resistant Staphylococcus aureus (MRSA). DESIGN. An unmatched case-control study (at a ratio of 1 : 2) performed during the period from October 2002 through September 2003. SETTING. A 2,000-bed tertiary care teaching hospital affiliated with the University of Sao Paulo in Sao Paulo, Brazil. METHODS. Case patients (n = 30) were defined either as patients who had a bloodstream infection due to SCCmec type IV strains of MRSA diagnosed at least 48 hours after hospital admission or as neonates with the infection who were born in the hospital. Control patients (n = 60) were defined as patients with SCCmec type III MRSA infection diagnosed at least 48 hours after hospital admission. Genes n = 60 encoding virulence factors were studied in the isolates recovered from case patients, and molecular typing of the SCCmec type IV MRSA isolates was also done by pulsed-field gel electrophoresis and multilocus sequence typing. RESULTS. In multivariate analysis, the following 3 variables were significantly associated with having a nosocomial bloodstream infection caused by SCCmec type IV strains of MRSA: an age of less than 1 year, less frequent use of a central venous catheter (odds ratio [OR], 0.07 [95% confidence interval {CI}, 0.02-0.28]; P = .001), and female sex. A second analysis was performed that excluded the case and Pp. 001 control patients from the neonatal unit, and, in multivariate analysis, the following variables were significantly associated with having a nosocomial bloodstream infection caused by SCCmec type IV strains of MRSA: less frequent use of a central venous catheter (OR, 0.12 [95% CI, 0.03-0.55]; P = .007), lower Acute Physiology and Chronic Health Evaluation II score on admission (OR, 0.14 [95% CI, 0.03-0.61];), less frequent surgery (OR, 0.21 [95% CI, 0.06-0.83];), and female sex (OR, 5.70 [95% CI, 1.32-24.66]; P =.020). P = .009 Pp. 025 Pp). Of the 29 SCCmec type IV MRSA isolates recovered from case patients, none contained the Panton-Valentine leukocidin, gamma-hemolysin, enterotoxin B or C, or toxic shock syndrome toxin-1. All of the isolates contained genes for the LukE-LukD leukocidin and alpha-hemolysin. Genes for enterotoxin A were present in 1 isolate, and genes for beta-hemolysin were present in 3 isolates. CONCLUSIONS. ""Classical"" risk factors do not apply to patients infected with the SCCmec type IV strain of MRSA, which is an important cause of nosocomial bacteremia. This strain infects a patient population that is less ill and has had less frequent invasive procedures than a patient population infected with the multidrug-resistant strain of SCCmec type III MRSA. We found that virulence factors were rare and that Panton-Valentine leukocidin was absent. There were multiple clones of the SCCmec type IV strain in our hospital. Children under 1 year of age were at a higher risk. There was a predominant clone ( sequence type 5) in this patient population.

Impact of antimicrobial resistance on the treatment and outcome of patients with sepsis

Antimicrobial therapy is one of the main stones of sepsis therapy. A recent study of septic shock patients showed that each hour of delay in antimicrobial administration during the ensuing 6 h after the onset of hypotension was associated with a decrease in survival rates. However, many questions regarding the impact of infection caused by antimicrobial-resistant pathogens on the mortality of patients with sepsis still need to be clarified. There is a lack of fair studies in the literature. Most studies have had inadequate sample size, inadequate adjustment for predictors of adverse outcomes, and inadequate definition of appropriate antibiotic therapy. Despite the fact that appropriate therapy is essential to treat sepsis, it seems that severity of underlying diseases and comorbidities are more important than resistance, although the studies were not well designed to examine the real impact of resistance on outcome. Finally, new technologies such as microarray that can identify different microorganisms, genes of resistance, and virulence in a few hours might have a great impact on the treatment of sepsis due to antimicrobial-resistant pathogens in the future.

New transposons to generate GFP protein fusions in Candida albicans

Transposon elements are important tools for gene function analysis, for example they can be used to easily create genome-wide collections of insertion mutants. Transposons may also carry sequences coding for an epitope or fluorescent marker useful for protein expression and localization analysis. We have developed three new Tn5-based transposons that incorporate a GFP (green fluorescent protein) coding sequence to generate fusion proteins in the important fungal pathogen Candida albicans. Each transposon also contains the URA3 and Kan(R) genes for yeast and bacterial selection, respectively. After in vitro transposition, the insertional allele is transferred to the chromosomal locus by homologous recombination. Transposons Tn5-CaGFP and Tn5-CaGFP-URA3:FLIP can generate C-terminal truncated GFP fusions. A URA3 flipper recycling cassette was incorporated into the transposon Th5-CaGFP-UFRA3:FLIP. After the induction of Flip recombinase to excise the marker, the heterozygous strain is transformed again in order to obtain a GFP-tagged homozygous strains. In the Tn5-CaGFP-FL transposon the markers are flanked by a rare-cutting enzyme. After in vitro transposition into a plasmid-borne target gene, the markers are eliminated by restriction digestion and religation, resulting in a construct coding for full-length GFP-fusion proteins. This transposon can generate plasmid libraries of GFP insertions in proteins where N- or C-terminal tagging may alter localization. We tested our transposon system by mutagenizing the essential septin CDC3 gene. The results indicate that the Cdc3 C-terminal extension is important for correct septin filament assembly. The transposons described here provide a new system to obtain global gene expression and protein localization data in C. albicans. (c) 2008 Elsevier B.V. All rights reserved.

Safety and Immunogenicity of Varicella Vaccine in Patients With Juvenile Rheumatic Diseases Receiving Methotrexate and Corticosteroids

Objective. To evaluate the safety and immunogenicity of varicella vaccine (VV) in susceptible patients with juvenile rheumatic diseases receiving methotrexate and corticosteroids. Methods. Twenty-five patients with juvenile rheumatic diseases (ages 2-19 years) and 18 healthy children and adolescents (ages 3-18 years) received a single dose of VV. All 25 patients were receiving methotrexate; 13 were also receiving prednisone and 5 were also receiving other disease-modifying antirheumatic drugs. None of the vaccinated patients or controls had a previous history of varicella. Anti-varicella-zoster virus IgG antibody (anti-VZV-IgG) titers were measured by enzyme-linked immunosorbent assay immediately before, 4-6 weeks after, and 1 year after vaccination. The patients were monitored prospectively for adverse reactions related to the vaccine, exposure, and occurrence of varicella. Disease activity was assessed 3 months before and 3 months after VV. Results. Twenty patients and all of the controls had negative preimmunization titers of VZV-IgG, and 5 patients had equivocal levels. Positive VZV-IgG titers were detected in 10 (50%) of 20 seronegative patients and 13 (72.2%) of 18 controls 4-6 weeks after VV (P = 0.2). One year after vaccination, 8 of 10 patients maintained positive VZV-IgG titers. No overt varicella episodes and no severe adverse reactions were observed during the followup period. No worsening of clinical parameters and no flares of juvenile rheumatic diseases or changes in doses of medications used were detected after vaccination. In fact, the number of active joints in patients with juvenile idiopathic arthritis was significantly lower after VV (P = 0.009). Conclusion. VV appears to be safe in patients with juvenile rheumatic diseases receiving methotrexate, as long as continuous prospective vigilance for side effects is performed.

Prevention of toothbrushing abrasion of acid-softened enamel by CO(2) laser irradiation

Objective: The aim of the present study was to evaluate the effect of CO(2) laser irradiation (10.6 mu m) at 0.3 J/cm(2) (0.5 mu s; 226 Hz) on the resistance of softened enamel to toothbrushing abrasion, in vitro. Methods: Sixty human enamel samples were obtained, polished with silicon carbide papers and randomly divided into five groups (n = 12), receiving 5 different surface treatments: laser irradiation (L), fluoride (AmF/NaF gel) application (F), laser prior to fluoride (LF), fluoride prior to laser (FL), non-treated control (C). After surface treatment they were submitted to a 25-day erosive-abrasive cycle in 100 ml sprite light (90 s) and brushed twice daily with an electric toothbrush. Between the demineralization periods samples were immersed in supersaturated mineral solution. At the end of the experiments enamel surface loss was determined using a contact profilometer and morphological analysis was performed using scanning electron microscopy (SEM). For SEM analysis of demineralization pattern, cross-sectional cuts of cycled samples were prepared. The data were statistically analysed by one-way ANOVA model with subsequent pairwise comparison of treatments. Results: Abrasive surface loss was significantly lower in all laser groups compared to both control and fluoride groups (p < 0.0001 in all cases). Amongst the laser groups no significant difference was observed. Softened enamel layer underneath lesions was less pronounced in laser-irradiated samples. Conclusion: Irradiation of dental enamel with a CO(2) laser at 0.3 J/cm(2) (5 mu s, 226 Hz) either alone or in combination with amine fluoride gel significantly decreases toothbrushing abrasion of softened-enamel, in vitro. (C) 2011 Elsevier Ltd. All rights reserved.

Rehardening of acid-softened enamel and prevention of enamel softening through CO(2) laser irradiation

Objectives: The aims of the present study were to investigate whether irradiation with a CO(2) laser could prevent surface softening (i) in sound and (ii) in already softened enamel in vitro. Methods: 130 human enamel samples were obtained and polished with silicon carbide papers. They were divided into 10 groups (n = 13) receiving 5 different surface treatments: laser irradiation (L), fluoride (AmF/NaF gel) application (F), laser prior to fluoride (LF), fluoride prior to laser (FL), non-treated control (C); and submitted to 2 different procedures: half of the groups was acid-softened before surface treatment and the other half after. Immersion in 1% citric acid was the acid challenge. Surface microhardness (SMH) was measured at baseline, after softening and after treatment. Additionally, fluoride uptake in the enamel was quantified. The data were statistically analysed by two-way repeated measurements ANOVA and post hoc comparisons at 5% significance level. Results: When softening was performed either before or after laser treatment, the L group presented at the end of the experiments SMH means that were not significantly different from baseline (p = 0.8432, p = 0.4620). Treatment after softening resulted for all laser groups in statistically significant increase in SMH means as compared to values after softening (p < 0.0001). Enamel fluoride uptake was significantly higher for combined laser-fluoride treatment than in control (p < 0.0001). Conclusion: Irradiation of dental enamel with a CO(2) laser at 0.3J/cm(2) (5 mu s, 226 Hz) not only significantly decreased erosive mineral loss (97%) but also rehardened previously softened enamel in vitro. (C) 2011 Elsevier Ltd. All rights reserved.

Direct comparison of the bond strength results of the different test methods: A critical literature review

Objective. The goal of this paper is to undertake a literature search collecting all dentin bond strength data obtained for six adhesives with four tests ( shear, microshear, tensile and microtensile) and to critically analyze the results with respect to average bond strength, coefficient of variation, mode of failure and product ranking. Method. A PubMed search was carried out for the years between 1998 and 2009 identifying publications on bond strength measurements of resin composite to dentin using four tests: shear, tensile, microshear and microtensile. The six adhesive resins were selected covering three step systems ( OptiBond FL, Scotch Bond Multi-Purpose Plus), two-step (Prime & Bond NT, Single Bond, Clear. l SE Bond) and one step (Adper Prompt L Pop). Results. Pooling results from 147 references showed an ongoing high scatter in the bond strength data regardless which adhesive and which bond test was used. Coefficients of variation remained high (20-50%) even with the microbond test. The reported modes of failure for all tests still included high number of cohesive failures. The ranking seemed to be dependant on the test used. Significance. The scatter in dentin bond strength data remains regardless which test is used confirming Finite Element Analysis predicting non-uniform stress distributions due to a number of geometrical, loading, material properties and specimens preparation variables. This reopens the question whether, an interfacial fracture mechanics approach to analyze the dentin - adhesive bond is not more appropriate for obtaining better agreement among dentin bond related papers. (C) 2009 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.