Yersinia enterocolitica YopH-Deficient Strain Activates Neutrophil Recruitment to Peyer's Patches and Promotes Clearance of the Virulent Strain.

Yersinia enterocolitica evades the immune response by injecting Yersinia outer proteins (Yops) into the cytosol of host cells. YopH is a tyrosine phosphatase critical for Yersinia virulence. However, the mucosal immune mechanisms subverted by YopH during in vivo orogastric infection with Y. enterocolitica remain elusive. The results of this study revealed neutrophil recruitment to Peyer's patches (PP) after infection with a YopH-deficient mutant strain (Y. enterocolitica ΔyopH). While the Y. enterocolitica wild-type (WT) strain in PP induced the major neutrophil chemoattractant CXCL1 mRNA and protein levels, infection with the Y. enterocolitica ΔyopH mutant strain exhibited a higher expression of the CXCL1 receptor, CXCR2, in blood neutrophils, leading to efficient neutrophil recruitment to the PP. In contrast, migration of neutrophils into PP was impaired upon infection with Y. enterocolitica WT strain. In vitro infection of blood neutrophils revealed the involvement of YopH in CXCR2 expression. Depletion of neutrophils during Y. enterocolitica ΔyopH infection raised the bacterial load in PP. Moreover, the clearance of WT Y. enterocolitica was improved when an equal mixture of Y. enterocolitica WT and Y. enterocolitica ΔyopH strains was used in infecting the mice. This study indicates that Y. enterocolitica prevents early neutrophil recruitment in the intestine and that the effector protein YopH plays an important role in the immune evasion mechanism. The findings highlight the potential use of the Y. enterocolitica YopH-deficient strain as an oral vaccine carrier.

Nitrogen sources differentially affect respiration, growth, and carbon allocation in Andean and Lowland ecotypes of Chenopodium quinoa Willd.

Chenopodium quinoa Willd. is a native species that originated in the High Andes plateau (Altiplano) and its cultivation spread out to the south of Chile. Because of the different edaphoclimatic characteristics of both regions, soils from Altiplano accumulated higher levels of nitrate (NO3-) than in the south of Chile, where soils favor ammonium (NH4 +) accumulation. To elucidate whether C. quinoa ecotypes differ in several physiological and biochemical parameters related to their capacity to assimilate NO3- and NH4 +, juvenile plants of Socaire (from Altiplano) and Faro (from Lowland/South of Chile) were grown under different sources of N (NO3- or NH4 +). Measurements of photosynthesis and foliar oxygen-isotope fractionation were carried out, together with biochemical analyses, as proxies for the analysis of plant performance or sensitivity to NH4 +. Overall, while NH4 + reduced the growth of Socaire, it induced higher biomass productivity and increased protein synthesis, oxygen consumption, and cytochrome oxidase activity in Faro. We discussed that ATP yield from respiration in Faro could promote protein production from assimilated NH4 + to benefit its growth. The characterization of this differential sensitivity of both quinoa ecotypes for NH4 + contributes to a better understanding of nutritional aspects driving plant primary productivity.

Procyanidins from grape pomace are suitable inhibitors of human endothelial NADPH oxidase.

Procyanidins have been associated with a reduced risk of cardiovascular diseases such as atherosclerosis. However, the molecular mechanisms underlying this benefit are not fully understood. Increased reactive oxygen species (ROS) production generated by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is a common problem in different cardiovascular diseases. Our objective was to evaluate the effects of procyanidin-rich fractions from distilled grape pomace on NADPH oxidase activity in human umbilical vein endothelial cells (HUVEC). Three differently polymerized and galloylated procyanidin fractions were analyzed for their NADPH oxidase inhibitory activity in cell lysates and in HUVEC cultures. All of the three fractions, up to 1 µg/ml, equally inhibited isolated NADPH oxidase in HUVEC lysates in a concentration-dependent manner and independently of any superoxide anion scavenging activities. The procyanidin fractions even blocked NADPH oxidase activity in intact HUVEC, inhibiting ROS production at both extra- and intracellular levels. The fractions achieved the same effects that known NADPH oxidase inhibitors, such as diphenylene iodonium and apocynin, but they presented better hydrosolubility. Our results demonstrated that procyanidin from grape pomace inhibit human endothelial NADPH oxidase regardless of their polymerization degree and galloylation percentage. Therefore, procyanidins are suitable NADPH oxidase inhibitors which could serve as models for therapeutic alternatives for cardiovascular diseases.

Massive bilateral grade IV emphysematous pyelonephritis: Case report.

Emphysematous pyelonephritis (EPN) is a severe disease of the renal system in which gas formed by facultative anaerobe microorganisms accumulates, being Escherichia coli the most representative causative agent. A series of conditions foster its development including uncontrolled diabetes mellitus, diabetic nephropathy and obstructive uropathy. Abdominal CT scan continues to be the gold standard for diagnosis. Currently, nephrectomy is avoided as much as possible, and more conservative treatments are given. Mortality is still as high as 21 % despite new therapeutic options, all of which are mostly surgical. The rarest cases are those classified as grade IV, which affect both kidneys; these are considered the most lethal of the clinical presentations and they are also of particular interest, since a multidisciplinary team must be in charge of treatment. Herein, we present the case of an elderly woman with chronic diseases and a presentation of massive emphysematous pyelonephritis concurrent with emphysematous cystitis and pneumoperitoneum, who required bilateral radical nephrectomy despite efforts of preserving at least one of the kidneys.

Trajectories of family and employment stress associated with cerebrovascular accidents.

OBJECTIVES: Reconstruct types of simultaneous stress trajectories in the family and employment domain at different stages of life and estimate their association with cerebrovascular accident (CVA) in old age. METHODS: We used a retrospective, face-to-face, representative survey of people aged 65 to 75 years in the city of Santiago, Chile, (n = 802). We performed a multichannel sequence analysis to reconstruct family and employment stress trajectory types at various life stages and then used logistic regression models to estimate the association of these trajectory types with CVA in old age, controlled for traditional cardiovascular risk factors. RESULTS: Four representative types of family and employment stress trajectories were identified: (1) Absence of family and employment stress, (2) Absence of family stress, persistent employment stress, (3) Absence of family stress, out of the labor market, and (4) Persistent family stress, absence of employment stress. The 61.7% of the sample followed trajectories marked by the permanent presence of family and/or employment stress. Likewise, 18.3% had a trajectory characterized by prolonged absence from the labor market. Individuals with persistent family or employment stress trajectories, as well as those with extended periods of inactivity, are more at risk of developing CVA. CONCLUSIONS: Stress is a risk factor for cardiovascular disease experienced by many people at different stages and domains of life on a prolonged basis. Consequently, prevention systems for this type of chronic diseases should emphasize the highly harmful effects of daily and cumulatively stressful life experiences. This could mitigate the multiple health and financial consequences associated with CVA.

Identification of polymeric procyanidins from pine bark by mass spectrometry.

Pine bark is an important source of polyphenolic compounds, mainly procyanidins, with reported protective effects against disease. In previous works, barks of two varieties of pine (P. pinaster and P. radiata) were extracted with ethanol, and partially purified to obtain the aqueous fractions (FA), that contained mainly polymeric procyanidins. The mean degree of polymerization was 7.9 for radiata (rFA) and 10.6 for pinaster (pFA). FAs were chromatographed on Sephadex LH-20 by using a gradient of methanol, water and acetone, to render a series of sub-fractions. In this work, the procyanidin compositions of these sub-fractions were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The mass spectra of sub-fractions from FA of P. pinaster showed signals of procyanidin polymers up to tridecamers, whereas for those from P. radiata the maximum degree of polymerization was 15. For this latter case, the MALDI-TOF mass spectra detected the presence of prodelphinidins in a small amount.

CODES, a novel procedure for ligand-based virtual screening: PDE7 inhibitors as an application example.

Phosphodiesterase (PDE) 7 is a high affinity cAMP-specific PDE whose functional role in T-cells has been the subject of some controversy. Recent findings on tissue distribution, however, support the hypothesis that PDE7 could be a good target for the treatment of airway diseases, T-cell related diseases or central nervous system (CNS) disorders. Therefore, the identification of selective inhibitors targeted against PDE7 enzyme has become an attractive area of research. We report here the first use of the descriptors generated by the CODES program for ligand-based virtual screening. This program codifies molecules from a topological point of view and the generated descriptors are related to the chemical nature of the atoms, the atomic bonds and the connectivity with the rest of the molecule. They are also able to distinguish among stereoisomers. By using this approach, 173 compounds were codified, and their similarity with the reference compound was analysed. Based on the analysis, new potential PDE7 inhibitors have been identified, synthesized and biologically evaluated confirming that CODES descriptors are valid for ligand-based virtual screening and provided new lead compounds for further optimization as potent and selective PDE7 inhibitors.

The activity of interleukin-4 receptor alpha-chain promoter is regulated by a GT box element.

Interleukin-4 receptor (IL-4R) is the cell surface complex through which interleukin-4 (IL-4) signals exert its critical biological effects. The alpha-chain of IL-4R is responsible for the high affinity binding of IL-4. In this report, is characterized, the 5' untranslated flanking region of murine IL-4Ralpha gene in the Th2 clone D10.G4.1. We have analyzed a DNA fragment spanning from -995 to +84 relative to the transcription start point. Mutagenesis analysis shows that, neither the previously described Stat6 (-395) nor the NFAT (-266) and NFkappaB (+25) sequences localized here, are involved in the IL-4Ralpha promoter activity. Reporter assays demonstrate that maximum transcriptional activity is achieved by the -89 to +84 sequence and this activity is independent of a TATA-like box located at -25. We have identified a GT box located at -45 as the critical element for the IL-4Ralpha promoter activity. Experiments in SL2 cells, which lack endogenous Sp proteins, show that IL-4Ralpha minimal promoter is transactivated by proteins of Sp family.

Trajectories of family and employment stress associated with cerebrovascular accidents / Trayectorias de estrés familiar y laboral y su asociación con accidentes cerebrovasculares

ABSTRACT OBJECTIVES: Reconstruct types of simultaneous stress trajectories in the family and employment domain at different stages of life and estimate their association with cerebrovascular accident (CVA) in old age. METHODS: We used a retrospective, face-to-face, representative survey of people aged 65 to 75 years in the city of Santiago, Chile, (n = 802). We performed a multichannel sequence analysis to reconstruct family and employment stress trajectory types at various life stages and then used logistic regression models to estimate the association of these trajectory types with CVA in old age, controlled for traditional cardiovascular risk factors. RESULTS: Four representative types of family and employment stress trajectories were identified: (1) Absence of family and employment stress, (2) Absence of family stress, persistent employment stress, (3) Absence of family stress, out of the labor market, and (4) Persistent family stress, absence of employment stress. The 61.7% of the sample followed trajectories marked by the permanent presence of family and/or employment stress. Likewise, 18.3% had a trajectory characterized by prolonged absence from the labor market. Individuals with persistent family or employment stress trajectories, as well as those with extended periods of inactivity, are more at risk of developing CVA. CONCLUSIONS: Stress is a risk factor for cardiovascular disease experienced by many people at different stages and domains of life on a prolonged basis. Consequently, prevention systems for this type of chronic diseases should emphasize the highly harmful effects of daily and cumulatively stressful life experiences. This could mitigate the multiple health and financial consequences associated with CVA.

RESUMEN OBJETIVOS: Reconstruir tipos de trayectorias de estrés simultáneo en el dominio familiar y laboral en diferentes etapas de la vida y estimar su asociación con accidentes cerebrovasculares (ACV) en la vejez. MÉTODOS: Se utilizó una encuesta retrospectiva, cara a cara, y representativa de personas de 65 a 75 años en la ciudad de Santiago de Chile (n = 802). Se empleó un análisis de secuencias multicanal para reconstruir tipos de trayectorias de estrés familiar y laboral en diversas etapas de la vida y luego se utilizaron modelos de regresión logística para estimar la asociación de estos tipos de trayectoria con ACV en la vejez, controlado por factores tradicionales de riesgo cardiovascular. RESULTADOS: Se identificaron cuatro tipos representativos de trayectorias de estrés familiar y laboral: (1) Ausencia de estrés familiar y laboral, (2) Ausencia de estrés familiar, estrés laboral persistente, (3) Ausencia de estrés familiar, fuera de mercado laboral, y (4) Estrés familiar persistente, ausencia de estrés laboral. El 61,7% de la muestra siguió trayectorias marcadas por la presencia permanente de estrés familiar y/o laboral. Asimismo, el 18,3% tuvo una trayectoria caracterizada por la ausencia prolongada del mercado del trabajo. Las personas con trayectorias de estrés familiar o laboral persistente, así como aquellas con períodos extensos de inactividad, tienen más riesgo de desarrollar ACV. CONCLUSIONES: El estrés es un factor de riesgo de enfermedades cardiovasculares que experimenta de manera prolongada muchas personas en distintas etapas y dominios de la vida. En consecuencia, los sistemas de prevención de este tipo de enfermedades crónicas deben enfatizar los efectos altamente nocivos de enfrentar cotidiana y acumulativamente experiencias de vida estresantes. Esto a su vez podría mitigar las múltiples consecuencias sanitarias y financieras asociadas al ACV.

Effect of solvent, temperature, and solvent-to-solid ratio on the total phenolic content and antiradical activity of extracts from different components of grape pomace.

Grape byproducts were subjected to an extraction process under various different experimental conditions (namely, solvent type, temperature, solvent-to-solid ratio, time contact, and raw material) in order to study the effect of these conditions on the yield of phenolic compounds and the corresponding antiradical activity of extracts. Although the order of decreasing capacity to extract soluble materials was ethanol > methanol > water, methanol was the most selective for extracting phenolic compounds. Temperature and solvent-to-solid ratio were found to have a critical role in extraction efficiency; values of 50 degrees C (between 25 and 50 degrees C) and 1:1 (between 1:1 and 5:1) maximized the antiradical activity of phenolic extracts. In addition, extracts from grape samples previously subjected to distillation reached higher antiradical values in comparison to those coming directly from pressing; in both cases, seed extracts showed better results than those of stem when ethanol or water was employed, whereas the opposite occurred in the case of methanol. These differences were attributed to the different phenolic compositions of the considered fractions.

Antiradical activity, phenolics profile, and hydroxymethylfurfural in espresso coffee: influence of technological factors.

The influence of technological factors (decaffeination, brew volume, coffee species, and roast degree) on antiradical activity and phenolics content of espresso coffee is described. The screenings of phenolics profile and other compounds (caffeine and trigonelline), as well as the quantification of hydroxymethylfurfural, were performed by LC-DAD-ESI-MS. Significantly lower (p < 0.05) scavenging activities and phenolics contents were found in decaffeinated espressos when compared with regular ones (32 vs 38% and 324 vs 410 mg/30 mL cup, respectively). A long espresso (70 mL) offers more than twice the phenolics amount of a short one (20 mL). Robusta brews showed higher (p < 0.05) antiradical activity and phenolic contents than arabica ones, for all roast degrees (light, medium, and dark). No significant differences (p > 0.05) were observed for scavenging activities of differently roasted robusta brews, whereas an increase in medium-dark brews was observed for arabica samples. Total phenolics in robusta espressos decreased (p < 0.05) with the increase of roast degree, but no significant differences (p > 0.05) were found between arabica espressos from different roasts. By LC-DAD-ESI-MS, 23 hydroxycinnamic derivatives were found, including chlorogenic acids, lactones, and cinnamoyl-amino acid conjugates. The amount of each compound was differently affected by species and roast. Robusta brews presented superior levels of caffeine and chlorogenic acids, whereas arabica ones contained more trigonelline. Hydroxymethylfurfural contents in the brew (30 mL) varied from 2.60 to 0.84 mg for light- and dark-roasted arabicas and from 1.29 to 0.68 mg for light- and dark-roasted robustas, respectively.

Confocal microscopy in ocular argyrosis.

PURPOSE: To identify the presence of silver deposits in the cornea using a new-generation confocal microscopy technique. METHODS: Case report. RESULTS: A 70-year-old man, who had been a jeweler for many years, was referred to our center for assessment of corneal opacity and ocular pigmentation. Slit-lamp examination revealed grayish, dense confluent deposits in the central and peripheral cornea, deep stroma, and Descemet's membrane. On confocal microscopy, we observed typical images of hyperreflective keratocytes across the entire stromal surface and two hyperreflective plaques coinciding with areas of metal deposition, one at Descemet's membrane and the other at Bowman's membrane. This last deposition site has not been previously identified in vivo by confocal microscopy. CONCLUSIONS: Confocal microscopy is a useful tool for the diagnosis of corneal argyrosis because it allows the in vivo visualization of silver deposits at different corneal levels.

Cyclic nucleotide phosphodiesterases and their role in immunomodulatory responses: advances in the development of specific phosphodiesterase inhibitors.

The activity of phosphodiesterases (PDEs) is associated with a wide variety of diseases and an intense effort toward the development of specific PDEs inhibitors has been generated for the last years. They are the enzymes responsible for the hydrolysis of intracellular cyclic adenosine and guanosine monophosphate, and their complexity, as well as their different functional role, makes these enzymes a very attractive therapeutic target. This review is focused on the role of PDEs played on immunomodulatory processes and the advance on the development of specific inhibitors, covering PDEs mainly related to the regulation of autoimmune processes, PDE4 and PDE7. The review also highlights the novel structural classes of PDE4 and PDE7 inhibitors, and the therapeutic potential that combined PDE4/PDE7 inhibitors offer as immunomodulatory agents.

Autocrine IL-4 gene regulation at late phases of TCR activation in differentiated Th2 cells.

IL-4 is a multifunctional cytokine whose secretion displays important immunomodulatory functions. Its expression is regulated at the level of transcription, and one of the main factors involved is NFAT. The IL-4-induced transcription factor Stat6 is required for the development of naive T cells into Th2 phenotype, capable of secreting IL-4. However, IL-4 production by differentiated Th2 cells is IL-4 independent; thus, it remains unclear whether Stat6 plays any role in the IL-4 expression by mature Th2 cells. We have analyzed in the Th2 clone D10.G4.1 the nuclear proteins able to bind the regulatory element P1 of the IL-4 promoter. Gel-shift assays show NFAT1 as the most abundant nuclear protein that binds to P1 after ionomycin plus PMA activation, whereas Stat6 accounts for the bulk of the P1 binding in the presence of exogenous IL-4. Reporter experiments agree with an inhibitory effect of Stat6 on the NFAT1-induced transcriptional activity directed by the P1 element. CD3 signaling leads to an early induction of NFAT1-P1 complexes correlating with a strong induction of the IL-4 gene. In later phases of CD3 activation, P1 is also bound by Stat6 and a fall in the IL-4 mRNA levels takes place. These two late events during CD3 activation were found to be sensible in experiments conducted with an anti-IL-4 Ab. These results suggest that IL-4 endogenously produced by Th2 cells under TCR triggering modulates its own expression through Stat6.

Hidranencefalia: caso clínico / Hydranencephaly: a clinical case

La hidranencefalia pertenece a las encefalopatías del sistema nervioso central. Es un defecto grave de la perfusión cerebral que produce una destrucción tardía de los territorios irrigados por ambas arterias carótidas internas, con preservación de los territorios irrigados por las arterias vertebro basilares. El conocimiento de esta encefalopatía por el odontopediatra permitirá tratar adecuadamente al paciente y realizar un tratamiento exitoso, mejorando su calidad de vida. Se presenta el caso clínico de un niño al que durante el quinto mes de vida intrauterina le fue realizada una ecografía donde se diagnosticó la encefalopatía.