Functional and structural characterisation of RimL from Bacillus cereus, a new Nα-acetyltransferase of ribosomal proteins that was wrongly assigned as an aminoglycosyltransferase.

Enzymes of the GNAT (GCN5-relate N-acetyltransferases) superfamily are important regulators of cell growth and development. They are functionally diverse and share low amino acid sequence identity, making functional annotation difficult. In this study, we report the function and structure of a new ribosomal enzyme, Nα-acetyl transferase from Bacillus cereus (RimLBC), a protein that was previously wrongly annotated as an aminoglycosyltransferase. Firstly, extensive comparative amino acid sequence analyses suggested RimLBC belongs to a cluster of proteins mediating acetylation of the ribosomal protein L7/L12. To assess if this was the case, several well established substrates of aminoglycosyltransferases were screened. The results of these studies did not support an aminoglycoside acetylating function for RimLBC. To gain further insight into RimLBC biological role, a series of studies that included MALDI-TOF, isothermal titration calorimetry, NMR, X-ray protein crystallography, and site-directed mutagenesis confirmed RimLBC affinity for Acetyl-CoA and that the ribosomal protein L7/L12 is a substrate of RimLBC. Last, we advance a mechanistic model of RimLBC mode of recognition of its protein substrates. Taken together, our studies confirmed RimLBC as a new ribosomal Nα-acetyltransferase and provide structural and functional insights into substrate recognition by Nα-acetyltransferases and protein acetylation in bacteria.

Socioeconomic Level and the Relationship in Glycemic Behavior in the Mexican Population. A Nutritional Alternative Focused on Vulnerable Populations.

In this study, numerical approximations were generated to analyze the behavior of glycemic variations in the non-diabetic population of the Mexican republic. The main objective of this work is to obtain an overview of the glycemic variations in the non-diabetic population from different socioeconomic statuses in Mexico (Medium-high, medium, and low). Thus, evaluating the effect on the glucose level under a healthy diet considering the socioeconomic capabilities of the population. Through the national health and nutrition survey of Mexico 2020 and the Mexican food base, 1420 virtual patients were proposed (522 low status, 485 medium status and 413 Medium-High status) focused on simulating the glycemic behavior in each of the survey participants. Considering that the average food expenditure of the Mexican population is $107.00 MXN, and the cost of a healthy diet is $66.50 MXN, the economic sustainability of the Mexican population to adopt a healthy diet is revealed. The particularity of this work is focused on obtaining diverse data that are difficult to access in the development of population analyses. Such is the case of the approach proposed for different socioeconomic statuses. In this way, the proposed methodology provides a framework for complementary research contributions to the subject.

Thermodynamic Study of Leaching Conditions of Galena with Citrate Ions and Hydrogen Peroxide as Oxidizing Agent.

Galena is the most important mineral for lead production, as it is the main source of lead in the world. Currently, the concentrates of this mineral are mainly treated using pyrometallurgical methods, creating several environmental problems, such as the generation of toxic and greenhouse gases. In addition, these processes involve high energy consumption, which limits their applicability. Hydrometallurgical routes are proposed as alternative processes for obtaining some metals such as silver, copper, gold, etc. The drawback of these processes is that the minerals tend to be passive in aqueous media. To mitigate this issue, researchers have used extreme conditions of pressure and temperature (6 atm. and 155 °C) or the use of very corrosive conditions. In this sense, the use of complexing agents that dissolve the metals of interest has been proposed. Citrate ion is one of the most promising complexing agents for galena leaching, obtaining high percentages of dissolution in relatively short times. Unfortunately, there has not been enough investigation about the concentration optimization of the complexing in the pH range from 5 to 9. In this sense, thermodynamic diagrams, such as the Pourbaix diagrams, are very useful for this purpose. Therefore, in this work, the effects of pH and temperature on the leaching of galena in citrate ion solutions are studied thermodynamically and experimentally. The experimental work was carried out with pure galena samples with a particle size of +149 - 74 µm (-100 + 200 mesh). The results show that higher recoveries were obtained working at a pH of 8 and at temperatures of 30 and 40 °C. The thermodynamic and experimental data demonstrated that the existence of an optimal concentration of citrate ion, due the extraction of lead from galena, has a greater reaction rate at a relatively low initial concentration of 0.3 M. This is due the formation of the complex lead citrate 1 (Pb(cit)-).

Photoinduced Dynamics with Constrained Vibrational Motion: FrozeNM Algorithm.

Ab initio molecular dynamics (AIMD) simulation, analyzed in terms of vibrational normal modes, is a widely used technique that facilitates understanding of complex structural motions and coupling between electronic and nuclear degrees of freedom. Usually, only a subset of vibrations is directly involved in the process of interest. The impact of these vibrations can be evaluated by performing AIMD simulations by selectively freezing certain motions. Herein, we present frozen normal mode (FrozeNM), a new algorithm to apply normal-mode constraints in AIMD simulations, as implemented in the nonadiabatic excited state molecular dynamics code. We further illustrate its capacity by analyzing the impact of normal-mode constraints on the photoinduced energy transfer between polyphenylene ethynylene dendrimer building blocks. Our results show that the electronic relaxation can be significantly slowed down by freezing a well-selected small subset of active normal modes characterized by their contributions in the direction of energy transfer. The application of these constraints reduces the nonadiabatic coupling between electronic excited states during the entire dynamical simulations. Furthermore, we validate reduced dimensionality models by freezing all the vibrations, except a few active modes. Altogether, we consider FrozeNM as a useful tool that can be broadly used to underpin the role of vibrational motion in a studied process and to formulate reduced models that describe essential physical phenomena.

Vibrational energy redistribution during donor-acceptor electronic energy transfer: criteria to identify subsets of active normal modes.

Photoinduced electronic energy transfer in conjugated donor-acceptor systems is naturally accompanied by intramolecular vibrational energy redistributions accepting an excess of electronic energy. Herein, we simulate these processes in a covalently linked donor-acceptor molecular dyad system by using nonadiabatic excited state molecular dynamics simulations. We analyze different complementary criteria to systematically identify the subset of vibrational normal modes that actively participate on the donor → acceptor (S2→ S1) electronic relaxation. We analyze energy transfer coordinates in terms of state-specific normal modes defined according to the different potential energy surfaces (PESs) involved. On one hand, we identify those vibrations that contribute the most to the direction of the main driving force on the nuclei during electronic transitions, represented by the non-adiabatic derivative coupling vector between donor and acceptor electronic states. On the other hand, we monitor normal mode transient accumulations of excess energy and their intramolecular energy redistribution fluxes. We observe that the subset of active modes varies according to the PES on which they belong and these modes experience the most significant rearrangements and mixing. Whereas the nuclear motions that promote donor → acceptor energy funneling can be localized mainly on one or two normal modes of the S2 state, they become spread out across multiple normal modes of the S1 state following the energy transfer event.

Nigrostriatal dopamine transporter availability, and its metabolic and clinical correlates in Parkinson's disease patients with impulse control disorders.

PURPOSE: Previous studies in patients with Parkinson's disease (PD) and impulse control disorders (ICDs) have produced heterogeneous results regarding striatal dopamine transporter (DaT) binding and activity in the mesocorticolimbic network. Our aim here was to study the relationship between striatal DaT availability and cortical metabolism, as well as motor, behavioural and cognitive features of PD patients with ICD. METHODS: In a group of PD patients with ICD (PD-ICD, n = 16) and 16 matched PD patients without ICD (PD-noICD, n = 16), DaT single-photon emission computed tomography (SPECT) imaging (DaTSCAN) was used to study DaT availability in predefined striatal volumes of interest (VOIs): putamen, caudate nucleus and ventral striatum (VS). In addition, the specific association of striatal DaT binding with cortical limbic and associative metabolic activity was evaluated by 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) in PD-ICD patients and investigated using statistical parametric mapping (SPM8). Finally, associations between DaT availability and motor, behavioural and cognitive features were assessed. RESULTS: PD-ICD patients had a significantly lower DaT density in the VS than PD-noICD patients, which was inversely associated with ICD severity. Lower DaT availability in the VS was associated with lower FDG uptake in several cortical areas belonging to the limbic and associative circuits, and in other regions involved in reward and inhibition processes (p < 0.0001 uncorrected; k > 50 voxels). No significant results were observed using a higher conservative threshold (p < 0.05; FDR corrected). PD-ICD patients also displayed impairment in interference and attentional Stroop Task execution, and more anxiety, all associated with reduced DaT availability in the VS and caudate nucleus. CONCLUSIONS: ICDs in PD patients are related to reduced DaT binding in the VS, which accounts for dysfunction in a complex cortico-subcortical network that involves areas of the mesolimbic and mesocortical systems, being associated with reward evaluation, salience attribution and inhibitory control processes.

[Estimation of the population attributable fraction due to obesity in hospital admissions for flu valued according to Body Mass Index (BMI) and CUN-BAE]. / Estimación de la fracción atribuible poblacional debida a la obesidad en los ingresos hospitalarios por gripe valorada según el índice de masa corporal (IMC) y el CUN-BAE.

INTRODUCTION: The obesity pandemic together with the influenza pandemic could lead to a significant burden of disease. The body mass index (BMI) does not discriminate obesity appropriately. The CUN-BAE has recently been used as an estimate of body fatness for Caucasians, including BMI, gender, and age. The aim of this study is to assess the population attributable fraction of hospital admissions due to influenza, due to the body fatness measured with the BMI, and the CUN-BAE. METHODS: A multicentre study was conducted using matched case-controls. Cases were hospital admissions with the influenza confirmed by the RT-PCR method between 2009 and 2011. The risk of hospital admission and the population attribuible fraction were calculated using the BMI or the CUN-BAE for each adiposity category in a conditional logical regression analysis adjusted for confounding variables. The analyzes were estimated in the total sample, in unvaccinated people, and those less than 65 years-old. RESULTS: A total of 472 hospitalised cases and 493 controls were included in the study. Compared to normal weight, the aOR of influenza hospital admissions increases with each level of BMI (aOR=1.26; 2.06 and 11.64) and CUN-BAE (aOR=2.78; 4.29; 5.43 and 15.18). The population attributable fraction of influenza admissions using CUN-BAE is 3 times higher than that estimated with BMI (0,72 vs. 0,27), with the differences found being similar the non-vaccinated and under 65 year-olds. CONCLUSION: The BMI could be underestimating the burden of disease attributable to obesity in individuals hospitalised with influenza. There needs to be an appropriate assessment of the impact of obesity and vaccine recommendation criteria.

Motor associations of iron accumulation in deep grey matter nuclei in Parkinson's disease: a cross-sectional study of iron-related magnetic resonance imaging susceptibility.

BACKGROUND AND PURPOSE: To determine whether iron deposition in deep brain nuclei assessed using high-pass filtered phase imaging plays a role in motor disease severity in Parkinson's disease (PD). METHODS: Seventy patients with mild to moderate PD and 20 age- and gender-matched healthy volunteers (HVs) underwent susceptibility-weighted imaging on a 3 T magnetic resonance imaging scanner. Phase shifts (radians) in deep brain nuclei were derived from high-pass filtered phase images and compared between groups. Analysis of clinical laterality and correlations with motor severity (Unified Parkinson's Disease Rating Scale, Part III, UPDRS-III) were performed. Phase shifts (in radians) were compared between HVs and three PD subgroups divided according to UPDRS-III scores using analysis of covariance, adjusting for age and regional area. RESULTS: Parkinson's disease patients had significantly (P < 0.001) higher radians than HVs bilaterally in the putamen, globus pallidus and substantia nigra (SN). The SN contralateral to the most affected side showed higher radians (P < 0.001) compared to the less affected side. SN radians positively correlated with UPDRS-III and bradykinesia-rigidity subscores, but not with tremor subscores. ancova followed by post hoc Bonferroni-adjusted pairwise comparisons revealed that SN radians were significantly greater in the PD subgroup with higher UPDRS-III scores compared to both lowest UPDRS-III PD and HV groups (P < 0.001). CONCLUSIONS: Increased nigral iron accumulation in PD appears to be stratified according to disease motor severity and correlates with symptoms related to dopaminergic neurodegeneration. This semi-quantitative in vivo iron assessment could prove useful for objectively monitoring PD progression, especially in clinical trials concerning iron chelation therapies.

High-Resolution Rotational Spectrum, Dunham Coefficients, and Potential Energy Function of NaCl.

We report laboratory spectroscopy for the first time of the J = 1-0 and J = 2-1 lines of Na35Cl and Na37Cl in several vibrational states. The hyperfine structure has been resolved in both transitions for all vibrational levels, which permit us to predict with high accuracy the hyperfine splitting of the rotational transitions of the two isotopologues at higher frequencies. The new data have been merged with all previous works at microwave, millimeter, and infrared wavelengths and fitted to a series of mass-independent Dunham parameters and to a potential energy function. The obtained parameters have been used to compute a new dipole moment function, from which the dipole moment for infrared transitions up to Δv = 8 has been derived. Frequency and intensity predictions are provided for all rovibrational transitions up to J = 150 and v = 8, from which the ALMA data of evolved stars can be modeled and interpreted.

Hints of a rotating spiral structure in the innermost regions around IRC +10216.

The Atacama Large Millimeter/submillimeter Array (ALMA) is allowing us to study the innermost regions of the circumstellar envelopes of evolved stars with un-precedented precision and sensitivity. Key processes in the ejection of matter and dust from these objects occur in their inner zones. In this work, we present sub-arcsecond interferometric maps of transitions of metal-bearing molecules towards the prototypical C-rich evolved star IRC +10216. While Al-bearing molecules seem to be present as a roughly spherical shell, the molecular emission from the salts NaCl and KCl presents an elongation in the inner regions, with a central minimum. In order to accurately analyze the emission from the NaCl rotational lines, we present new calculations of the collisional rates for this molecule based on new spectroscopic constants. The most plausible interpretation for the spatial distribution of the salts is a spiral with a NaCl mass of 0.08M☉. Alternatively, a torus of gas and dust would result in similar structures as those observed. From the torus scenario we derive a mass of ~ 1.1 × 10-4M☉. In both cases, the spiral and the torus, the NaCl structure presents an inner minimum of 27 AU. In the case of the torus, the outer radius is 73 AU. The kinematics of both the spiral and the torus suggests that they are slowly expanding and rotating. Alternative explanations for the presence of the elongation are explored. The presence of these features only in KCl and NaCl might be a result of their comparatively high dipole moment with respect to the Al-bearing species.

SI-BEARING MOLECULES TOWARD IRC+10216: ALMA UNVEILS THE MOLECULAR ENVELOPE OF CWLEO.

We report the detection of SiS rotational lines in high-vibrational states as well as SiO and SiC2 lines in their ground vibrational state toward IRC+10216 during the Atacama Large Millimeter Array Cycle 0. The spatial distribution of these molecules shows compact emission for SiS and a more extended emission for SiO and SiC2, and also proves the existence of an increase in the SiC2 emission at the outer shells of the circumstellar envelope. We analyze the excitation conditions of the vibrationally excited SiS using the population diagram technique, and we use a large velocity gradient model to compare with the observations. We found moderate discrepancies between the observations and the models that could be explained if SiS lines detected are optically thick. Additionally, the line profiles of the detected rotational lines in the high energy vibrational states show a decreasing linewidth with increasing energy levels. This may be evidence that these lines could be excited only in the inner shells, i.e., the densest and hottest, of the circumstellar envelope of IRC+10216.

A theoretical study of the reaction of beta-carotene with the nitrogen dioxide radical in solution.

A theoretical study of the reaction of beta-carotene (BC) with the nitrogen dioxide radical (NO2*) in solution is carried out using the density functional theory (DFT) at the B3LYP/6-31G(d) level, to optimize the molecular geometries, and the polarizable continuum model (PCM), to account for solvent effects. The three most important reaction mechanisms--electron transfer from beta-carotene to the radical, hydrogen abstraction by the radical, and radical addition to form an adduct--are studied in detail. Three solvents with different polarities--heptane, methanol, and water--are employed to investigate the effect of the environment on the reaction mechanisms. Our results show that electron transfer is thermodynamically favored only in the polar solvents, the abstraction reactions are spontaneous in the three solvents, although faster in the polar ones, and the addition reactions are all endergonic and, therefore, unlikely to occur in any of the solvents. In both the abstraction and addition mechanisms, the attack of the radical takes place preferentially at the beta-ionone rings, in particular at positions H4 and C5, respectively. The higher stability of the reaction products in these cases is explained in terms of their molecular geometries and electronic structures.

Density functional theory study of the stability and vibrational spectra of the beta-carotene isomers.

A density functional theory analysis of the stability and vibrational spectra of the beta-carotene isomers is carried out. The study includes the 7-, 9-, 11-, 13-, and 15-monocis isomers and the 7,13'-, 9,13-, 9,13'-, 9,15-, 11,11'-, and 13,15-dicis isomers. The optimized geometries needed to study the stability of the isomers are calculated at the B3LYP/6-31G(d) level of theory, and their energies are further recalculated at the higher B3LYP/6-311+G(2d,2p) level. In addition, the Wiberg bond orders and the natural bond orbital charges of the isomers are computed study the effect of the torsion of the beta-ionone rings on the conjugation degree of the polyene chain. The infrared and Raman spectra of the beta-carotene isomers are then calculated at the B3LYP/6-31G(d) level, scaling the calculated frequencies with an overall factor to account for the anharmonicity effects. The calculated frequencies are shown to compare quite well with the experiment, and the normal modes of the key bands are theoretically interpreted.

A density functional theory study of the structure and vibrational spectra of beta-carotene, capsanthin, and capsorubin.

A theoretical study of the structure and the vibrational spectra of the beta-carotene molecule and its derivatives capsanthin and capsorubin is carried out. We first investigate systematically the theoretical method which provides the best results for beta-carotene by performing ab initio calculations at the HF/6-31G(d), SVWN/6-31G(d), PBE0/6-31G(d), BLYP/6-31G(d), B3LYP/6-31G(d), B3LYP/6-31G(d,p), B3LYP/6-311G(d), and B3LYP/6-311G(d,p) levels and by using previous theoretical results available in the literature obtained at the AM1 and BPW91/6-31G(d) levels. The influence of both the level of calculation and the size of the basis set used in the geometry optimization and in the determination of the IR and Raman spectra of this molecule is thus analyzed. It is confirmed that the hybrid functional B3LYP with the basis 6-31G(d) is the method that gives the best results as a whole. By use of this level of calculation, we next optimize the molecular geometries of related molecules of capsanthin and capsorubin, which to the best of our knowledge have only been studied at the semiempirical AM1 level. In addition we calculate the IR and Raman spectra of these molecules at the B3LYP/6-31G(d) level of theory. The results obtained for capsanthin show on the one hand that the double bond of the beta-ionone ring is outside the polyene chain plane, due to the repulsion between the hydrogen atoms of the ring methyl groups and the hydrogen atoms of the polyene chain, and on the other hand that the carbonyl double bond in the other headgroup is very close to planarity with the polyene chain, since in this case such a repulsion does not exist. For the molecule of capsorubin the two carbonyl groups also take the same coplanar orientation relative to the polyene chain. The IR and Raman spectra theoretically computed for these two molecules are finally compared with their experimental spectra and the vibrational normal modes of the main signals are interpreted.

[Written information on the use of aerosols in COPD patients. Can we improve their use?]. / Información escrita sobre el uso de aerosoles en pacientes con enfermedad pulmonar obstructiva crónica. Mejoramos su calidad de utilización?

OBJECTIVES: To determine whether the use of written information alone improved inhalation technique with pressurized canister inhalers in patients with chronic obstructive pulmonary disease (COPD). To compare the effectiveness of written information versus verbal explanation provided by nursing personnel on the use of inhalers. DESIGN: Interventional study. SETTING: Urban health center. PARTICIPANTS: 120 patients with COPD treated with pressurized canister inhalers. INTERVENTIONS: The participants were divided randomly into three groups of 40 patients each. In one group no intervention was used, in the second group verbal explanations were provided, and in the third group written information was provided. MAIN MEASURES: We recorded percentage compliance with 5 criteria for the correct use of inhalers at the start of the study and 3 months after the intervention in all groups. RESULTS: Initially, performance of the inhalation technique by patients with COPD was poor (mean compliance 40%). Performance improved significantly in both intervention groups, with no significant difference between them. Final mean compliance was 74% in the written information group and 82% in the verbal information group. CONCLUSIONS: The use of written information about the use of inhalers for patients with COPD significantly improved utilization to a degree similar to that obtained with verbal explanations.

Size evolution of the vibrational predissociation process in Br2...Nen clusters: simulation and kinetic study.

A hybrid quantum-classical simulation of the vibrational predissociation of Br2...Nen, (n = 2-11) clusters in the B electronic state is carried out. The time-evolution of the reactants, products, and intermediates is analyzed by a kinetic mechanism consisting of three elementary steps: direct vibrational predissociation (VP), intramolecular vibrational redistribution (IVR), and evaporative cooling (EC). The importance of intramolecular vibrational redistribution followed by evaporative cooling relative to direct vibrational predissociation is shown to increase rapidly with increasing cluster size. Final product state distributions reveal that only one or less Br2 stretching quantum per neon atom is required in order to achieve complete dissociation (n quanta for n < or = 9 and n - 1 for n = 10 and 11). The proportion of available energy going into translation is proposed as a parameter to study the statistical behavior of the Van der Waals clusters. It is shown to depend only on the number of remaining degrees of freedom, a characteristic of a statistical behavior, for n > or = 3.

Heterologous over-expression of alpha-1,6-fucosyltransferase from Rhizobium sp.: application to the synthesis of the trisaccharide beta-D-GlcNAc(1-->4)-[alpha-L-Fuc-(1-->6)]-D-GLcNAc, study of the acceptor specificity and evaluation of polyhydroxylated indolizidines as inhibitors.

An efficient heterologous expression system for overproduction of the enzyme alpha-1,6-Fucosyltransferase (alpha-1,6-FucT) from Rhizobium sp. has been developed. The gene codifying for the alpha-1,6-FucT was amplified by PCR using specific primers. After purification, the gene was cloned in the plasmid pKK223-3. The resulting plasmid, pKK1,6FucT, was transformed into the E. coli strain XL1-Blue MRF'. The protein was expressed both as inclusion bodies and in soluble form. Changing the induction time a five-fold increase of enzyme expressed in soluble form was obtained. In this way five units of enzyme alpha-1,6-FucT can be obtained per liter of culture. A crude preparation of the recombinant enzyme was used for the synthesis of the branched trisaccharide alpha-D-GlcNAc-(1-->4)-[alpha-L-Fuc-(1-->6)]-D-GlcNAc (3), from chitobiose (2) and GDP-Fucose (1). After purification, the trisaccharide 3 was obtained in a 84% overall yield. In order to elucidate the structural requirements for the acceptors, the specificity of the enzyme was studied towards mono-, di- and trisaccharides, which are structurally related to chitobiose. The enzyme uses, among others, the disaccharide N-acetyl lactosamine as a good substrate; the monosaccharide GlcNAc is a weak acceptor. Finally, several racemic polyhydroxylated indolizidines have been tested as potential inhibitors of the enzyme. Indolizidine 21 was the best inhibitor with an IC50 of 4.5 x 10(-5) M. Interestingly, this compound turned out to be the best mimic for the structural features of the fucose moiety in the presumed transition state.