Plasma corticosterone elevation inhibits the activation of nuclear factor kappa B (NFKB) in the Syrian hamster pineal gland

We evaluated how the mild stress-induced increase in endogenous corticosterone affected the pineal gland in Syrian hamsters (Mesocricetus auratus). The animals were maintained under constant light for 1 day, instead of a cycle of 14:10-h, to increase the circulating corticosterone levels during the daytime. The nuclear translocation of nuclear factor kappa B (NFKB), which is the pivotal transcription factor for stress and injury, presented a daily rhythm in normal animals. NFKB nuclear content increased linearly from the onset of light [Zeitgeber Time 0 (ZT0)] until ZT11 and decreased after ZT12 when the plasma corticosterone peak was detected in normal animals. However, the 24-h profiles of the two curves were different, and they did not clearly support an exclusive relationship between corticosterone levels and NFKB content. Therefore, we tested the effect of increased endogenous corticosterone through inducing mild stress by maintaining daytime illumination for one night. This stressful condition, which increased daytime corticosterone levels, resulted in a daytime decrease in NFKB nuclear content, and this was inhibited by mifepristone. Overall, this study shows that NFKB has a daily rhythm in Syrian hamster pineal glands and, by increasing endogenous corticosterone with a stressful condition, NFKB activity is regulated. Therefore, this study suggests that the pineal gland in the Syrian hamster is a sensor of stressful conditions.

Partitioning and extraction of collagenase from Penicillium aurantiogriseum in poly(ethylene glycol)/phosphate aqueous two-phase system

Purification of collagenase produced by Penicillium aurantiogriseum URM4622 was carried using a PEG/phosphate aqueous two-phase system (ATPS). A 2(3)-full experimental design was used to investigate the influence of PEG molar mass, PEG concentration and phosphate concentration on the selected responses, namely partition coefficient, activity yield and purification factor. The ATPS was composed of PEG (molar mass of 550, 1500 and 4000 g/mol) at concentrations of 15.0, 17.5 and 20.0% (w/w) and phosphate at concentrations of 12.5, 15.0 and 17.5% (w/w). The best results of one-step extraction of collagenase from the fermentation broth (partition coefficient of 1.01, activity yield of 242% and purification factor of 23.5) were obtained at pH 6.0 using 20.0% (w/w) PEG 550 and 17.5% (w/w) phosphate. The results of this preliminary study demonstrate that the selected ATPS is satisfactorily selective for the extraction of such a collagenase. (C) 2012 Elsevier B.V. All rights reserved.

IRF6 is a Risk Factor for Nonsyndromic Cleft Lip in the Brazilian Population

Nonsyndromic cleft lip with or without cleft palate (NSCL/P) is a complex disorder with a worldwide incidence estimated at 1:700. Among the putative susceptibility loci, the IRF6 gene and a region at 8q24.21 have been corroborated in different populations. To test the role of IRF6 in NSCL/P predisposition in the Brazilian population, we conducted a structured association study with the SNPs rs642961 and rs590223, respectively, located at 5' and 3' of the IRF6 gene and not in strong linkage disequilibrium (LD), in patients from five different Brazilian locations. We also evaluated the effect of these SNPs in IRF6 expression in mesenchymal stem cells (MSC). We observed association between rs642961 and cleft lip only (CLO) (P = 0.009; odds ratio (OR) for AA genotype = 1.83 [95% Confidence interval (CI), 0.64-5.31]; OR for AG genotype = 1.72 [95% CI, 1.03-2.84]). This association seems to be driven by the affected patients from Barbalha, a location which presents the highest heritability estimate (H-2 = 0.85), and the A allele at rs642961 is acting through a dominant model. No association was detected for the SNP rs590223. We did not find any correlation between expression levels and genotypes of the two loci, and it is possible that these SNPs have a functional role in some specific period of embryogenesis. (C) 2012 Wiley Periodicals, Inc.

Extracytoplasmic function (ECF) sigma factor sigma(F) is involved in Caulobacter crescentus response to heavy metal stress

Background: The alpha-proteobacterium Caulobacter crescentus inhabits low-nutrient environments and can tolerate certain levels of heavy metals in these sites. It has been reported that C. crescentus responds to exposure to various heavy metals by altering the expression of a large number of genes. Results: In this work, we show that the ECF sigma factor sigma(F) is one of the regulatory proteins involved in the control of the transcriptional response to chromium and cadmium. Microarray experiments indicate that sigma(F) controls eight genes during chromium stress, most of which were previously described as induced by heavy metals. Surprisingly, sigma(F) itself is not strongly auto-regulated under metal stress conditions. Interestingly, sigma(F)-dependent genes are not induced in the presence of agents that generate reactive oxygen species. Promoter analyses revealed that a conserved sigma(F)-dependent sequence is located upstream of all genes of the sigma(F) regulon. In addition, we show that the second gene in the sigF operon acts as a negative regulator of sigma(F) function, and the encoded protein has been named NrsF (Negative regulator of sigma F). Substitution of two conserved cysteine residues (C131 and C181) in NrsF affects its ability to maintain the expression of sigma(F)-dependent genes at basal levels. Furthermore, we show that sigma(F) is released into the cytoplasm during chromium stress and in cells carrying point mutations in both conserved cysteines of the protein NrsF. Conclusion: A possible mechanism for induction of the sigma(F)-dependent genes by chromium and cadmium is the inactivation of the putative anti-sigma factor NrsF, leading to the release of sigma(F) to bind RNA polymerase core and drive transcription of its regulon.

Possible Host Adaptation as an Evolution Factor of Cowpea aphid-borne mosaic virus Deduced by Coat Protein Gene Analysis

Cowpea aphid-borne mosaic virus (CABMV) causes major diseases in cowpea and passion flower plants in Brazil and also in other countries. CABMV has also been isolated from leguminous species including, Cassia hoffmannseggii, Canavalia rosea, Crotalaria juncea and Arachis hypogaea in Brazil. The virus seems to be adapted to two distinct families, the Passifloraceae and Fabaceae. Aiming to identify CABMV and elucidate a possible host adaptation of this virus species, isolates from cowpea, passion flower and C.hoffmannseggii collected in the states of Pernambuco and Rio Grande do Norte were analysed by sequencing the complete coat protein genes. A phylogenetic tree was constructed based on the obtained sequences and those available in public databases. Major Brazilian isolates from passion flower, independently of the geographical distances among them, were grouped in three different clusters. The possible host adaptation was also observed in fabaceous-infecting CABMV Brazilian isolates. These host adaptations possibly occurred independently within Brazil, so all these clusters belong to a bigger Brazilian cluster. Nevertheless, African passion flower or cowpea-infecting isolates formed totally different clusters. These results showed that host adaptation could be one factor for CABMV evolution, although geographical isolation is a stronger factor.

Features of two proteins of Leptospira interrogans with potential role in host-pathogen interactions

Background: Leptospirosis is considered a re-emerging infectious disease caused by pathogenic spirochaetes of the genus Leptospira. Pathogenic leptospires have the ability to survive and disseminate to multiple organs after penetrating the host. Leptospires were shown to express surface proteins that interact with the extracellular matrix (ECM) and to plasminogen (PLG). This study examined the interaction of two putative leptospiral proteins with laminin, collagen Type I, collagen Type IV, cellular fibronectin, plasma fibronectin, PLG, factor H and C4bp. Results: We show that two leptospiral proteins encoded by LIC11834 and LIC12253 genes interact with laminin in a dose - dependent and saturable mode, with dissociation equilibrium constants (K-D) of 367.5 and 415.4 nM, respectively. These proteins were named Lsa33 and Lsa25 (Leptospiral surface adhesin) for LIC11834 and LIC12253, respectively. Metaperiodate - treated laminin reduced Lsa25 - laminin interaction, suggesting that sugar moieties of this ligand participate in this interaction. The Lsa33 is also PLG - binding receptor, with a K-D of 23.53 nM, capable of generating plasmin in the presence of an activator. Although in a weak manner, both proteins interact with C4bp, a regulator of complement classical route. In silico analysis together with proteinase K and immunoflorescence data suggest that these proteins might be surface exposed. Moreover, the recombinant proteins partially inhibited leptospiral adherence to immobilized laminin and PLG. Conclusions: We believe that these multifunctional proteins have the potential to participate in the interaction of leptospires to hosts by mediating adhesion and by helping the bacteria to escape the immune system and to overcome tissue barriers. To our knowledge, Lsa33 is the first leptospiral protein described to date with the capability of binding laminin, PLG and C4bp in vitro.

Performance tests of two small trigeneration pilot plants

Trigeneration systems have been used with advantage in the last years in distributed electricity generation systems as a function of a growth of natural gas pipeline network distribution system, tax incentives, and energy regulation policies. Typically, a trigeneration system is used to produce electrical power simultaneously with supplying heating and cooling load by recovering the combustion products thermal power content that otherwise would be driven to atmosphere. Concerning that, two small scale trigeneration plants have been tested for overall efficiency evaluation and operational comparison. The first system is based on a 30 kW (ISO) natural gas powered microturbine, and the second one uses a 26 kW natural gas powered internal combustion engine coupled to an electrical generator as a prime mover. The stack gases from both machines were directed to a 17.6 kW ammonia-water absorption refrigeration chiller for producing chilled water first and next to a water heat recovery boiler in order to produce hot water. Experimental results are presented along with relevant system operational parameters for appropriate operation including natural gas consumption, net electrical and thermal power production, i.e., hot and cold water production rates, primary energy saving index, and the energy utilization factor over total and partial electrical load operational conditions. (c) 2011 Elsevier Ltd. All rights reserved.

Caffeic acid phenethyl ester reduces the activation of the nuclear factor kappa B pathway by high-fat diet-induced obesity in mice

Objective. The aim of this study was to investigate the effect of CAPE on the insulin signaling and inflammatory pathway in the liver of mice with high fat diet induced obesity. Material/Methods. Swiss mice were fed with standard chow or high-fat diet for 12-week. After the eighth week, animals in the HFD group with serum glucose levels higher than 200 mg/dL were divided into two groups, HFD and HFD receiving 30 mg/kg of CAPE for 4 weeks. After 12 weeks, the blood samples could be collected and liver tissue extracted for hormonal and biochemical measurements, and insulin signaling and inflammatory pathway analyzes. Results. The high-fat diet group exhibited more weight gain, glucose intolerance, and hepatic steatosis compared with standard diet group. The CAPE treatment showed improvement in glucose sensitivity characterized by an area under glucose curve similar to the control group in an oral glucose tolerance test Furthermore, CAPE treatment promoted amelioration in hepatic steatosis compared with the high-fat diet group. The increase in glucose sensitivity was associated with the improvement in insulin-stimulated phosphorylation of the insulin receptor substrate-2, followed by an increase in Akt phosphorylation. In addition, it was observed that CAPE reduced the induction of the inflammatory pathway, c-jun-N- terminal kinase, the nuclear factor kappa B, and cyclooxygenase-2 expression, respectively. Conclusions. Overall, these findings indicate that CAPE exhibited anti-inflammatory activity that partly restores normal metabolism, reduces the molecular changes observed in obesity and insulin resistance, and therefore has a potential as a therapeutic agent in obesity. (C) 2012 Elsevier Inc. All rights reserved.

Effects of Two Different Levels of Dietary Protein on Body Composition and Protein Nutritional Status of Growing Rats

This study aimed to investigate the effect of a high-protein diet on growth, body composition, and protein nutritional status of young rats. Newly-weaned Wistar rats, weighing 45-50 g, were distributed in two experimental groups, according to their diets, which contained 12% (G12) or 26% protein (G26), over a period of 3 weeks. The animals were euthanized at the end of this period and the following analyses were performed: chemical composition of the carcass, proteoglycan synthesis, IGF-I concentration (serum, muscle and cartilage), total tissue RNA, protein concentration (muscle and cartilage) and protein synthesis (muscle and cartilage). The high-protein diet was found to result in a higher fat-free mass and lower fat mass in the carcass, with no difference in growth or protein nutritional status.

The effect of curing light and chemical catalyst on the degree of conversion of two dual cured resin luting cements

The aim of this study was to evaluate the influence of different curing lights and chemical catalysts on the degree of conversion of resin luting cements. A total of 60 disk-shaped specimens of RelyX ARC or Panavia F of diameter 5 mm and thickness 0.5 mm were prepared and the respective chemical catalyst (Scotchbond Multi-Purpose Plus or ED Primer) was added. The specimens were light-cured using different curing units (an argon ion laser, an LED or a quartz-tungsten-halogen light) through shade A2 composite disks of diameter 10 mm and thickness 2 mm. After 24 h of dry storage at 37A degrees C, the degree of conversion of the resin luting cements was measured by Fourier-transformed infrared spectroscopy. For statistical analysis, ANOVA and the Tukey test were used, with p a parts per thousand currency signaEuro parts per thousand 0.05. Panavia F when used without catalyst and cured using the LED or the argon ion laser showed degree of conversion values significantly lower than RelyX ARC, with and without catalyst, and cured with any of the light sources. Therefore, the degree of conversion of Panavia F with ED Primer cured with the quartz-tungsten-halogen light was significantly different from that of RelyX ARC regardless of the use of the chemical catalyst and light curing source. In conclusion, RelyX ARC can be cured satisfactorily with the argon ion laser, LED or quartz-tungsten-halogen light with or without a chemical catalyst. To obtain a satisfactory degree of conversion, Panavia F luting cement should be used with ED Primer and cured with halogen light.

No-pole condition in Landau gauge: Properties of the Gribov ghost form factor and a constraint on the 2d gluon propagator

We study general properties of the Landau-gauge Gribov ghost form factor sigma(p(2)) for SU(N-c) Yang-Mills theories in the d-dimensional case. We find a qualitatively different behavior for d = 3, 4 with respect to the d = 2 case. In particular, considering any (sufficiently regular) gluon propagator D(p(2)) and the one-loop-corrected ghost propagator, we prove in the 2d case that the function sigma(p(2)) blows up in the infrared limit p -> 0 as -D(0) ln(p(2)). Thus, for d = 2, the no-pole condition sigma(p(2)) < 1 (for p(2) > 0) can be satisfied only if the gluon propagator vanishes at zero momentum, that is, D(0) = 0. On the contrary, in d = 3 and 4, sigma(p(2)) is finite also if D(0) > 0. The same results are obtained by evaluating the ghost propagator G(p(2)) explicitly at one loop, using fitting forms for D(p(2)) that describe well the numerical data of the gluon propagator in two, three and four space-time dimensions in the SU(2) case. These evaluations also show that, if one considers the coupling constant g(2) as a free parameter, the ghost propagator admits a one-parameter family of behaviors (labeled by g(2)), in agreement with previous works by Boucaud et al. In this case the condition sigma(0) <= 1 implies g(2) <= g(c)(2), where g(c)(2) is a "critical" value. Moreover, a freelike ghost propagator in the infrared limit is obtained for any value of g(2) smaller than g(c)(2), while for g(2) = g(c)(2) one finds an infrared-enhanced ghost propagator. Finally, we analyze the Dyson-Schwinger equation for sigma(p(2)) and show that, for infrared-finite ghost-gluon vertices, one can bound the ghost form factor sigma(p(2)). Using these bounds we find again that only in the d = 2 case does one need to impose D(0) = 0 in order to satisfy the no-pole condition. The d = 2 result is also supported by an analysis of the Dyson-Schwinger equation using a spectral representation for the ghost propagator. Thus, if the no-pole condition is imposed, solving the d = 2 Dyson-Schwinger equations cannot lead to a massive behavior for the gluon propagator. These results apply to any Gribov copy inside the so-called first Gribov horizon; i.e., the 2d result D(0) = 0 is not affected by Gribov noise. These findings are also in agreement with lattice data.

Effects of Hepatocyte Growth Factor Injection and Reinjection on Healing in the Rabbit Vocal Fold

Objectives/Hypothesis. Hepatocyte growth factor (HGF) is a multifunctional polypeptide that plays various roles in embryogenesis and tissue regeneration and exhibits marked antifibrotic activity. The present study sought to assess the effects of HGF injection and reinjection coinciding with its peak of activity on collagen density, vessel density, inflammatory reaction in the lamina propria, and mean epithelial thickness in the injured rabbit vocal fold. Study Design. Prospective, controlled, experimental animal study. Methods. Fourteen rabbits were subdivided into two groups and underwent injury of the vocal folds. Immediately after injury, animals in group 1 received HGF injections into the right vocal fold (RVF), whereas those in group 2 received bilateral HGF injections and a single reinjection into the RVF 10 days after the first, to coincide with the peak of HGF activity. The left vocal folds (LVFs) served as controls in both groups. Histological assessment of laryngeal specimens was performed at 30 and 40 days, respectively. Results. In both groups, collagen density was lower in the right (treated) vocal folds than in the left (control) folds (P = 0.018). Vessel density was higher in the RVFs in group 2 (P = 0.018). Differences were found in mean epithelial thickness and inflammatory reaction in the lamina propria but did not reach statistical significance. Conclusions. In the scarred rabbit vocal fold, HGF injection is associated with decreased collagen density in the lamina propria, whereas reinjection after 10 days produces decreased collagen density and higher vessel density.

Comparison of two experimental models of pulmonary hypertension

Objective: To compare two models of pulmonary hypertension (monocrotaline and monocrotaline+pneumonectomy) regarding hemodynamic severity, structure of pulmonary arteries, inflammatory markers (IL-1 and PDGF), and 45-day survival. Methods: We used 80 Sprague-Dawley rats in two study protocols: structural analysis; and survival analysis. The rats were divided into four groups: control; monocrotaline (M), pneumonectomy (P), and monocrotaline+pneumonectomy (M+P). In the structural analysis protocol, 40 rats (10/group) were catheterized for the determination of hemodynamic variables, followed by euthanasia for the removal of heart and lung tissue. The right ventricle (RV) was dissected from the interventricular septum (IS), and the ratio between RV weight and the weight of the left ventricle (LV) plus IS (RV/LV+IS) was taken as the index of RV hypertrophy. In lung tissues, we performed histological analyses, as well as using ELISA to determine IL-1 and PDGF levels. In the survival protocol, 40 animals (10/group) were followed for 45 days. Results: The M and M+P rats developed pulmonary hypertension, whereas the control and P rats did not. The RV/LV+IS ratio was significantly higher in M+P rats than in M rats, as well as being significantly higher in M and M+P rats than in control and P rats. There were no significant differences between the M and M+P rats regarding the area of the medial layer of the pulmonary arteries; IL-1 and PDGF levels; or survival. Conclusions: On the basis of our results, we cannot conclude that the monocrotaline+pneumonectomy model is superior to the monocrotaline model.

Crowding: risk factor or protective factor for lower respiratory disease in young children?

Abstract Background To study the effects of household crowding upon the respiratory health of young children living in the city of São Paulo, Brazil. Methods Case-control study with children aged from 2 to 59 months living within the boundaries of the city of São Paulo. Cases were children recruited from 5 public hospitals in central São Paulo with an acute episode of lower respiratory disease. Children were classified into the following diagnostic categories: acute bronchitis, acute bronchiolitis, pneumonia, asthma, post-bronchiolitis wheezing and wheezing of uncertain aetiology. One control, crudely matched to each case with regard to age (<2, 2 years old or more), was selected among healthy children living in the neighborhood of the case. All buildings were surveyed for the presence of environmental contaminants, type of construction and building material. Plans of all homes, including measurements of floor area, height of walls, windows and solar orientation, was performed. Data were analysed using conditional logistic regression. Results A total of 313 pairs of children were studied. Over 70% of the cases had a primary or an associated diagnosis of a wheezing illness. Compared with controls, cases tended to live in smaller houses with less adequate sewage disposal. Cases and controls were similar with respect to the number of people and the number of children under five living in the household, as well the number of people sharing the child's bedroom. After controlling for potential confounders, no evidence of an association between number of persons sharing the child's bedroom and lower respiratory disease was identified when all cases were compared with their controls. However, when two categories of cases were distinguished (infections, asthma) and each category compared separately with their controls, crowding appeared to be associated with a 60% reduction in the incidence of asthma but with 2 1/2-fold increase in the incidence of lower respiratory tract infections (p = 0.001). Conclusion Our findings suggest that household crowding places young children at risk of acute lower respiratory infection but may protect against asthma. This result is consistent with the hygiene hypothesis.

Toll-like receptor 2 knockdown modulates Interleukin (IL)-6 and IL-8 but not Stromal Derived Factor-1 (SDF-1/CXCL12) in human periodontal ligament and gingival fibroblasts

Background: Fibroblasts are now seen as active components of the immune response because these cells express Toll-like receptors (TLRs), recognize pathogen-associated molecular patterns, and mediate the production of cytokines and chemokines during inflammation. The innate host response to lipopolysaccharide (LPS) from Porphyromonas gingivalis is unusual inasmuch as different studies have reported that it can be an agonist for Toll-like receptor 2 (TLR2) and an antagonist or agonist for Toll-like receptor 4 (TLR4). This study investigates and compares whether signaling through TLR2 or TLR4 could affect the secretion of interleukin (IL)-6, IL-8, and stromal derived factor-1 (SDF-1/CXCL12) in both human gingival fibroblasts (HGF) and human periodontal ligament fibroblasts (HPDLF). Methods: After small interfering RNA-mediated silencing of TLR2 and TLR4, HGF and HPDLF from the same donors were stimulated with P. gingivalis LPS or with two synthetic ligands of TLR2, Pam2CSK4 and Pam3CSK4, for 6 hours. IL-6, IL-8, and CXCL12 mRNA expression and protein secretion were evaluated by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Results: TLR2 mRNA expression was upregulated in HGF but not in HPDLF by all the stimuli applied. Knockdown of TLR2 decreased IL-6 and IL-8 in response to P. gingivalis LPS, or Pam2CSK4 and Pam3CSK4, in a similar manner in both fibroblasts subpopulations. Conversely, CXCL12 remained unchanged by TLR2 or TLR4 silencing. Conclusion: These results suggest that signaling through TLR2 by gingival and periodontal ligament fibroblasts can control the secretion of IL-6 and IL-8, which contribute to periodontal pathogenesis, but do not interfere with CXCL12 levels, an important chemokine in the repair process.