Unveiling the Shape of N-Acetylgalactosamine: A Cancer-Associated Sugar Derivative.

In the present work, we report the first rotational study of N-acetylgalactosamine, a cancer-associated sugar derivative, by means of high-resolution rotational spectroscopy. Two different conformers have been conclusively characterized using broadband Fourier transform microwave spectroscopy coupled with a laser ablation vaporization system. Additionally, we performed a comprehensive analysis of the intramolecular interactions that govern these structures, which allowed us to both characterize the existence of intramolecular hydrogen bond networks that drive the intrinsic conformation panorama of N-acetylgalactosamine and further rationalize the biological role of this aminosugar derivative as part of the Tn antigen.

Understanding the abundance of the rare sugar ß-D-allose.

The conformational landscape of ß-D-allose, a rare sugar, was investigated using laser ablation in combination with high-resolution rotational spectroscopy. Altogether, three species are identified, exhibiting a counter-clockwise intramolecular hydrogen bond network. The effect of epimerization on the main aldohexose is also studied and, despite the main conformers being very similar, the position of the hydroxyl groups in allose allows the formation of considerably stronger intramolecular hydrogen bonds than in glucose, and this could explain the low abundance of ß-D-allose in Nature.

The eight structures of caffeic acid: a jet-cooled laser ablated rotational study.

This work reports a complete conformational analysis of caffeic acid, an exceptionally versatile pharmacophore, using laser ablation chirped-pulse Fourier transform microwave spectroscopy. The whole conformational space consisting of eight distinct species has been fully deciphered based on the trend of the rotational constants supported by theoretical computations. We show how rotational spectroscopy can be confidently used to distinguish between conformers even when the structural differences are minimal, such as those involved in the conformational panorama of caffeic acid. Additionally, the structural information here provided, such as the planarity observed in all the conformers, could help to elucidate the mechanisms underlying the biological and pharmacological activity of hydroxycinnamic acids.

Interstellar glycolamide: A comprehensive rotational study and an astronomical search in Sgr B2(N).

CONTEXT: Glycolamide is a glycine isomer and also one of the simplest derivatives of acetamide (e.g., one hydrogen atom is replaced with a hydroxyl group), which is a known interstellar molecule. AIMS: In this context, the aim of our work is to provide direct experimental frequencies of the ground vibrational state of glycolamide in the centimeter-, millimeter- and submillimeter-wavelength regions in order to enable its identification in the interstellar medium. METHODS: We employed a battery of state-of-the-art rotational spectroscopic techniques in the frequency and time domain to measure the frequencies of glycolamide. We used the spectral line survey named Exploring Molecular Complexity with ALMA (EMoCA), which was performed toward the star forming region Sgr B2(N) with ALMA to search for glycolamide in space. We also searched for glycolamide toward Sgr B2(N) with the Effelsberg radio telescope. The astronomical spectra were analyzed under the local thermodynamic equilibrium approximation. We used the gas-grain chemical kinetics model MAGICKAL to interpret the results of the astronomical observations. RESULTS: About 1500 transitions have been newly assigned up to 460 GHz to the most stable conformer, and a precise set of spectroscopic constants was determined. Spectral features of glycolamide were then searched for in the prominent hot molecular core Sgr B2(N2). We report the nondetection of glycolamide toward this source with an abundance at least six and five times lower than that of acetamide and glycolaldehyde, respectively. Our astrochemical model suggests that glycolamide may be present in this source at a level just below the upper limit, which was derived from the EMoCA survey. We could also not detect the molecule in the region's extended molecular envelope, which was probed with the Effelsberg telescope. We find an upper limit to its column density that is similar to the column densities obtained earlier for acetamide and glycolaldehyde with the Green Bank Telescope.

Formation of interstellar cyanoacetamide: a rotational and computational study.

CONTEXT: Cyanoacetamide is a -CN bearing molecule that is also an amide derivative target molecule in the interstellar medium. AIMS: The aim of our investigation is to analyze the feasibility of a plausible formation process of protonated cyanoacetamide under interstellar conditions and to provide direct experimental frequencies of the ground vibrational state of the neutral form in the microwave region in order to enable its eventual identification in the interstellar medium. METHODS: We used high-level theoretical computations to study the formation process of protonated cyanoacetamide. Furthermore, we employed a high-resolution laser-ablation molecular beam Fourier transform spectroscopic technique to measure the frequencies of the neutral form. RESULTS: We report the first rotational characterization of cyanoacetamide, and a precise set of the relevant rotational spectroscopic constants have been determined as a first step to identifying the molecule in the interstellar medium. We fully explored the potential energy surface to study a gas-phase reaction on the formation process of protonated cyanoacetamide. We found that an exothermic process with no net activation barrier is initiated by the high-energy isomer of protonated hydroxylamine, which leads to protonated cyanoacetamide.

Association between obesity and mortality in adult patients receiving invasive mechanical ventilation: A systematic review and meta-analysis. / Asociación entre obesidad y mortalidad en pacientes adultos que reciben ventilación mecánica invasiva: una revisión sistemática y metaanálisis.

OBJECTIVE: To evaluate if there is an association between obesity and mortality in the Intensive Care Unit (ICU) in adult patients receiving invasive mechanical ventilation. DESIGN: Systematic review with meta-analysis. SCOPE: ICU. DATA SOURCE: A search was made in MEDLINE, Cochrane Library, CINAHL and Global Health databases without language restriction, until February 21, 2017. SELECTION OF STUDIES: Studies that reported mortality in the ICU in obese versus non-obese patients who received IMV were included. MAIN VARIABLES: Mortality in the ICU. RESULTS: 2163 articles were found, of which 14 studies were included. No statistically significant differences were found between obese and non-obese patients with respect to the variable mortality in the ICU (OR: 0.94, 95% CI: 0.81-1.10, P=.45). CONCLUSION: No relationship was found between the subgroup of obese adult patients receiving IMV and the mortality variable in the ICU.

Applying sunscreens on earthworms: Molecular response of Eisenia fetida after direct contact with an organic UV filter.

The use of organic Ultraviolet (UV) filters has increased in the last years, either in sunscreens, other cosmetics, or even food packaging. These filters may end up in soil and water since the Wastewater Treatment Plants may not successfully remove them. Among them, benzophenones are known to act as endocrine disruptors. However, most of the studies are directed towards vertebrates and aquatic invertebrates, while there is a lack of information on the molecular mechanisms affected by these compounds on soil dwelling invertebrates. Here, we study the impact of direct acute (48 h) contact of 4-hydroxybenzophenone (4-OHBP) at two sublethal concentrations (0.02 and 0.2 mg/mL) on gene expression of the earthworm Eisenia fetida. Investigated genes were involved in endocrine pathways, stress response, detoxification mechanisms, genotoxicity, energy metabolism and epigenetics. Three of them were identified for the first time in earthworms. Our results suggest that exposure to 4-OHBP affected endocrine pathways, causing an increase in the Ecdysone receptor gene (EcR) expression. Moreover, the UV filter induced changes in the CuZn superoxide dismutase gene (CuZn SOD), indicating an effect in the stress response. Finally, significant changes were detected for glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) expression, indicating that energy metabolism is influenced by the 4-OHBP and highlighting the risks of using GAPDH as an internal reference for Real Time PCR.

Bisphenol A in artificial soil: Effects on growth, reproduction and immunity in earthworms.

The application of biosolids in agricultural fields is increasing annually. They contain not only nutrients but also xenobiotics, such as Bisphenol A (BPA). These compounds are not regulated in the use of biosolids in agriculture, which highlights the need to assess their effects on soil life, of which earthworms are most abundant of the animal representatives. In this study the effect of BPA on life-history parameters, such as mortality, growth and reproduction, and on immunity, is evaluated for Dendrobaena veneta and Eisenia fetida. Sublethal concentrations were evaluated by a modified OECD artificial soil test. Decline in growth with increasing concentration of BPA was detected during the first two weeks and the opposite effect for the next two, although these differences were only significant at the highest concentration. Reproduction traits were only significantly different for E. fetida, for which the number of juveniles decreased at higher concentrations, thus showing different sensitivity in both species. By using a contact test, the potentially harmful effect of direct contact with BPA was shown to be much higher than in soil (resembling natural) conditions. Finally, results indicate that BPA may not affect the immune system of these animals, at least in terms of coelomocyte viability.

Endocrine disruptors in soil: Effects of bisphenol A on gene expression of the earthworm Eisenia fetida.

Xenobiotics such as bisphenol A (BPA), are present in biosolids, which are applied as organic fertilizers in agricultural fields. Their effects on soil life have been poorly assessed, and this is particularly important in the case of earthworms, which represent the main animal biomass in this medium. In the present work we study the impacts of BPA on gene expression of Eisenia fetida, a widely used ecotoxicological model. Chronic soil tests and acute contact tests were performed, and gene expression was analyzed in total tissue and in masculine reproductive organs of the earthworms. The genes studied in this research played a role in endocrine pathways, detoxification mechanisms, stress response, epigenetics, and genotoxicity. Most of the genes were identified for the first time, providing potentially useful biomarkers for future assessments. For chronic exposures, no changes were detected in whole-body tissue; however, masculine reproductive organs showed changes in the expression of genes related to endocrine function (EcR, MAPR, AdipoR), epigenetic mechanisms (DNMTs), genotoxicity (PARP1), and stress responses (HSC70 4). For acute exposures, the expression of one epigenetic-related gene was altered for both whole-body tissues and male reproductive organs (Piwi2). Further changes were detected for whole-body tissues involved in detoxification (Metallothionein), stress (HSC70 4), and genotoxicity (PARP1) mechanisms. Acute exposure effects were also tested in whole-body tissues of juveniles, showing changes in the expression of Metallothionein and Piwi2. The molecular changes found in the analyzed earthworms indicate that exposure to BPA may have negative implications in their populations. Particularly interesting are the alterations related to epigenetic mechanisms, which suggest that future generations may be impacted. This study is the first to evaluate the molecular effects of BPA on soil organisms, and further assays will be necessary to better characterize the true environmental repercussions. CAPSULE: Levels of gene expression in total-body tissues and masculine reproductive organs were analyzed in earthworms after exposure to bisphenol A and we observed associated changes in detoxification, endocrine, epigenetic, genotoxic and stress pathways.

Low-grade endotoxemia, gut permeability and platelet activation in patients with impaired fasting glucose.

BACKGROUND AND AIM: Impaired fasting glucose (IFG) is associated with an increased risk of cardiovascular disease but the underlying mechanisms are still unclear. Aim of the study was to investigate the interplay between platelet activation, lipopolysaccharides (LPS) and markers of oxidative stress in patients with IFG and control subjects. METHODS AND RESULTS: We performed a cross-sectional study including 35 patients with IFG and 35 control subjects who were well comparable for age, sex, body mass index and smoking history. Serum levels of LPS, zonulin (a marker of gut permeability), oxidized LDL and plasma levels of soluble P-selectin, were measured. Patients with IFG had significantly higher levels of sP-selectin, LPS, zonulin and oxLDL compared to control subjects. The IFG status (beta coefficient: 0.518, p < 0.001), higher LPS (beta coefficient: 0.352, p = 0.001) and female sex (beta coefficient: 0.179, p = 0.042) were independently associated with higher sP-selectin; in addition, oxLDL was positively associated with sP-selectin (r = 0.530, p < 0.001) and LPS (r = 0.529, p = 0.001). In IFG patients, we found a significant association between LPS and zonulin (r = 0.521, p = 0.001); this association was confirmed at multivariable analysis (beta coefficient: 0.512, p = 0.007). CONCLUSION: Our study provides evidence that patients with IFG have increased platelet activation, and suggests LPS as a potential trigger for in vivo platelet activation in this patient population.

Effects of dark chocolate on endothelial function in patients with non-alcoholic steatohepatitis.

BACKGROUND AND AIM: Oxidative stress plays a pivotal role in inducing endothelial dysfunction and progression from simple fatty liver steatosis (FLD) to non-alcoholic steatohepatitis (NASH). Polyphenols could reduce oxidative stress and restore endothelial function by inhibiting the nicotinamide-adenine-dinucleotide-phosphate (NADPH) oxidase isoform Nox2. The aim of this study was to assess endothelial function and oxidative stress in a population affected by simple FLD and NASH. Furthermore, we analysed the effect of high vs low content of cocoa polyphenols on endothelial function and oxidative stress in patients with NASH. METHODS: In a cross-sectional study we analysed endothelial function, as assessed by flow-mediated dilation (FMD), and oxidative stress, as assessed by Nox2 activation, serum isoprostanes and nitric oxide bioavailability (NOx), in patients with NASH (n = 19), FLD (n = 19) and controls (n = 19). Then, we performed a randomized, cross-over study in 19 subjects with NASH comparing the effect of 14-days administration of 40 g of chocolate at high (dark chocolate, cocoa >85%) versus low content (milk chocolate, cocoa <35%) of polyphenols on FMD and oxidative stress. Compared to controls, NASH and FLD patients had higher Nox2 activity and isoprostanes levels and lower FMD and NOx, with a significant gradient between FLD and NASH. The interventional study showed that, compared to baseline, FMD and NOx increased (from 2.9 ± 2.4 to 7.2 ± 3.0% p < 0.001 and from 15.9 ± 3.6 to 20.6 ± 4.9 µM, p < 0.001, respectively) in subjects given dark but not in those given milk chocolate. A simple linear regression analysis showed that Δ (expressed by difference of values between before and after 14 days of chocolate assumption) of FMD was associated with Δ of Nox2 activity (Rs = -0.323; p = 0.04), serum isoprostanes (Rs: -0.553; p < 0.001) and NOx (Rs: 0.557; p < 0.001). CONCLUSIONS: Cocoa polyphenols improve endothelial function via Nox2 down-regulation in NASH patients.

Ventilación mecánica invasiva como tratamiento de la insuficiencia respiratoria aguda en pacientes infectados por el virus de la inmunodeficiencia humana / Invasive mechanical ventilation as a treatment of Severe respiratory insufficiency in patients infected by the immunodeficiency virus human

Objetivo: Conocer la prevalencia y las características epidemiológicas de los pacientes infectados por el virus de la inmunodeficiencia humana que requirieron ventilación mecánica invasiva más de 12 horas para tratar la insuficiencia respiratoria aguda en la Unidad de Cuidados Intensivos de un Hospital General de Agudos. Asimismo, se realizó un análisis comparativo entre los pacientes con el virus de la inmunodeficiencia humana y aquellos sin el virus. Materiales y Métodos: Estudio descriptivo, prospectivo y longitudinal llevado a cabo entre el 1 de agosto de 2012 y el 31 de julio de 2014. Se incluyeron adultos con ventilación mecánica invasiva por más de 12 horas debido a insuficiencia respiratoria aguda. Se realizó un análisis multivariado de regresión logística para identificar la asociación entre muerte en la Unidad de Cuidados Intensivos y virus de la inmunodeficiencia humana. Resultados: Ingresaron 344 pacientes en la Unidad, el 46,80% requirió ventilación mecánica invasiva por insuficiencia respiratoria aguda, con una prevalencia del virus de la inmunodeficiencia humana del 12,42%. Los pacientes infectados tenían una media de la edad de 39.42 ± 11.58 vs. 49.37 ± 20.54. En un análisis multivariado, se observó que los pacientes infectados corrían un mayor de riesgo de morir en la Unidad de Cuidados Intensivos que los no infectados (OR: 5,125; IC95% 1,725-15,226; p = 0,003). Conclusión: Los pacientes con el virus de la inmunodeficiencia humana que recibieron ventilación mecánica invasiva más de 12 horas para tratar la insuficiencia respiratoria aguda tuvieron un riesgo cinco veces más alto de morir en la Unidad de Cuidados Intensivos que los no infectados. (AU)

Objective: To know the prevalence and epidemiological characteristics of patients infected with human immunodeficiency virus requiring invasive mechanical ventilation for more than 12 hours as a treatment for acute respiratory failure at the Intensive Care Unit of a General Acute Care Hospital in Buenos Aires City. A comparative analysis was also performed between subjects with human immunodeficiency virus and those not infected. Materials and Methods: Descriptive, prospective and longitudinal study conducted between August 1st, 2012 and July 31st, 2014. Adults with invasive mechanical ventilation for more than 12 hours due to acute respiratory failure were included. A multivariate logistic regression analysis was performed to identify the association between mortality in the Intensive Care Unit and human immunodeficiency virus. Results: A total of 344 patients were admitted to the Intensive Care Unit, 46.80% of them required invasive mechanical ventilation for acute respiratory failure, with a prevalence of human immunodeficiency virus of 12.42%. The average age of infected patients was 39.42 ± 11.58 vs. 49.37 ± 20.54. In a multivariate analysis it was observed that patients with human immunodeficiency virus had more risk of death in the Intensive Care Unit than those not infected (OR: 5.12%, CI95% 1.72-15.22; P=0.003). Conclusion: The risk of death of subjects with human immunodeficiency virus who received invasive mechanical ventilation for more than 12 hours as a treatment for acute respiratory failure was five-fold higher than that of those uninfected patients.(AU)

Molecular detection of Wolbachia pipientis in natural populations of mosquito vectors of Dirofilaria immitis from continental Portugal: first detection in Culex theileri.

Wolbachia pipientis (Rickettsiales: Rickettsiaceae) protects mosquitoes from infections with arboviruses and parasites. However, the effect of its co-infection on vector competence for Dirofilaria immitis (Spirurida: Onchocercidae) in the wild has not been investigated. This study aimed to screen vectors of D. immitis for wPip, to characterize these, and to investigate a possible association between the occurrence of W. pipientis and that of the nematode. The presence of W. pipientis was assessed in the five mosquito potential vectors of D. immitis in Portugal. Polymerase chain reaction (PCR) products were sequenced, and wPip haplotypes were determined by PCR-restricted fragment length polymorphism (RFLP). Results showed that wPip was detected in 61.5% of Culex pipiens (Diptera: Culicidae) pools and 6.3% of Culex theileri pools. wPip 16s rRNA sequences found in Cx. theileri exactly match those from Cx. pipiens, confirming a mosquito origin, rather than a nematode origin, as some specimens were infected with D. immitis. Only wPip haplotype I was found. No association was found between the presence of wPip and D. immitis in mosquitoes and hence a role for this endosymbiont in influencing vectorial competence is yet to be identified. This study contributes to understanding of wPip distribution in mosquito populations and, to the best of the authors' knowledge, is the first report of natural infections by wPip in Cx. theileri.

[Difficult encounters in primary care: A multifocal perspective]. / Encuentros difíciles en atención primaria: una perspectiva multifocal.

INTRODUCTION: The main objective of the study is to analyze the perception of difficult encounters experienced by the various professional groups involved in patient care. MATERIAL AND METHODS: The methodology used is qualitative, using focus groups. Four groups were formed: doctors, nurses, primary care receptionists, and experts in communication skills. The location was primary care, in Ourense area. Sampling was purposive. Patients were recruited by a member of the same professional group. Each session was recorded on audio and video. Information was categorized by reference to the bibliography and information resulting from conversations. RESULTS: The following categories were established: the concept of a difficult patient/difficult encounter, reasons for difficulty, consequences for the professional and patient, individual strategies of management, and strategies that institutions should take. The concept of difficulty varies among the different groups. All are capable of recognizing the professional responsibility, but this aspect does not arise spontaneously, except in the group of experts. In relation to difficult encounter management, most express strategies based on common sense and experience. The expert group uses more sophisticated techniques. CONCLUSIONS: There are differences in the quality of discourse and positioning relative to the user from the group of experts and other groups. Training in communication techniques is recommended to encourage a change in attitudes towards a patient-centered code, but the content of training should be developed based on knowledge of the group to which they are addressed.

Living on a volcano's edge: genetic isolation of an extremophile terrestrial metazoan.

Communities of organisms inhabiting extreme terrestrial environments provide a unique opportunity to study evolutionary forces that drive population structure and genetic diversity under the combined challenges posed by multiple geogenic stressors. High abundance of an invasive pantropical earthworm (and the absence of indigenous lumbricid species) in the Furnas geothermal field (Sao Miguel Island, Azores) indicates its remarkable tolerance to high soil temperature, exceptionally high carbon dioxide and low oxygen levels, and elevated metal bioavailability, conditions which are lethal for the majority of terrestrial metazoans. Mitochondrial and nuclear markers were used to analyze the relationship between populations living inside and outside the geothermal field. Results showed that Pontoscolex corethrurus (Annelida, Oligochaeta, Glossoscolecidae) to be a genetically heterogeneous complex within the Sao Miguel landscape and is probably differentiated into cryptic species. The population exposed to the hostile soil conditions within the volcanic caldera possesses the lowest within-population mitochondrial diversity but an unexpectedly high degree of nuclear variability with several loci evidencing positive selection, parameters indicative of a genetically unique population only distantly related to conspecifics living outside the caldera. In conclusion, P. corethrurus inhabiting active volcanic soil is a discrete extremophile population that has evolved by tolerating a mixture of non-anthropogenic chemical and physical stressors.

Unemployment and ill health--a gender analysis: results from a 14-year follow-up of the Northern Swedish Cohort.

OBJECTIVES: To analyse the associations between unemployment and suboptimal self-rated health as well as high alcohol consumption, and to examine the role of possible mediating factors explaining the associations from a gender perspective. STUDY DESIGN: The sample, from a 14-year longitudinal study with a 96.4% response rate, consisted of 386 women and 478 men who were either employed or unemployed at 30 years of age. METHODS: The health outcomes studied were suboptimal self-rated health and high alcohol consumption at 30 years of age. Logistic regression was used for analysis, and the relational theory of gender was used to discuss the findings. RESULTS: A strong relationship was found between unemployment and suboptimal self-rated health among women, and unemployment and high alcohol consumption among men, even after controlling for health-related selection, potential mediators and background factors. All mediating factors in the model were attributable to suboptimal self-rated health among unemployed women. Two mediating factors were also substantially related to high alcohol consumption among unemployed men. CONCLUSIONS: Long-term unemployment at a young age could have various health effects in men and women. At present, the mechanisms behind the health consequences are better understood among women. Research would benefit from developing theories in order to explain how youth unemployment leads to gendered health consequences.

Recombinant expression and characterization of a cysteine peptidase from Xanthomonas citri subsp citri.

Xanthomonas citri subsp citri (Xac) is the bacterium responsible for citrus canker disease in citrus plants. The aim of this study was to describe the recombinant expression, purification, and characterization of a cysteine peptidase from Xac strain 306, which is a candidate for involvement in the pathogenicity of this bacterium. The gene was cloned and expressed in Pichia pastoris, and the cysteine peptidase was successfully expressed, secreted, and purified using affinity chromatography with a yield of approximately 10 mg/L. A polyclonal antibody produced against cysteine peptidase from X. citri subsp citri fused with HIS tag ((HIS)CPXAC) recognized the purified recombinant cysteine peptidase (HIS)CPXAC, confirming the correct production of this protein in P. pastoris. The same antibody detected the protein in the culture supernatant of Xac grown in pathogenicity-inducing medium. Kinetic analysis revealed that (HIS)CPXAC hydrolyzed the carbobenzoxy-Leu-Arg-7-amido-4-methylcoumarin substrate with a catalytic efficiency (k(cat)/K(m)) of 47 µM(-1)∙s(-1). The purified ((HIS))CPXAC displayed maximal catalytic activity at pH 5.5 and 30°C. The recombinant enzyme was inhibited by the specific cysteine peptidase inhibitor E-64, as well as by the recombinant cysteine peptidase inhibitors CaneCPI-1, CaneCPI-2, CaneCPI-3, and CaneCPI-4, with K(i) values of 1.214, 84.64, 0.09, 0.09, and 0.012 nM, respectively. Finally, the N-terminal sequencing of the purified protein enabled the identification of the first 5 amino acid residues (AVHGM) immediately after the putative signal peptide, thereby enabling the identification of the cleavage point and corroborating previous studies that have identified this sequence in a secreted protein from Xanthomonas spp.

Ongoing outbreak of dengue type 1 in the Autonomous Region of Madeira, Portugal: preliminary report.

Following the identification of two autochthonous cases of dengue type 1 on 3 October 2012, an outbreak of dengue fever has been reported in Madeira, Portugal. As of 25 November, 1,891 cases have been detected on the island where the vector Aedes aegypti had been established in some areas since 2005. This event represents the first epidemic of dengue fever in Europe since 1928 and concerted control measures have been initiated by local health authorities.