Identification of duplicated and stress-inducible Aox2b gene co-expressed with Aox1 in species of the Medicago genus reveals a regulation linked to gene rearrangement in leguminous genomes

In flowering plants, alternative oxidase (Aox) is encoded by 3-5 genes distributed in 2 subfamilies (Aox1 and Aox2). In several species only Aox1 is reported as a stress-responsive gene, but in the leguminous Vigna unguiculata Aox2b is also induced by stress. In this work we investigated the Aox genes from two leguminous species of the Medicago genus (Medicago sativa and Medicago truncatula) which present one Aox1, one Aox2a and an Aox2b duplication (named here Aox2b1 and Aox2b2). Expression analyses by semi-quantitative RT-PCR in M. sativa revealed that Aox1, Aox2b1 and Aox2b2 transcripts increased during seed germination. Similar analyses in leaves and roots under different treatments (SA, PEG, H2O2 and cysteine) revealed that these genes are also induced by stress, but with peculiar spatio-temporal differences. Aox1 and Aox2b1 showed basal levels of expression under control conditions and were induced by stress in leaves and roots. Aox2b2 presented a dual behavior, i.e., it was expressed only under stress conditions in leaves, and showed basal expression levels in roots that were induced by stress. Moreover, Aox2a was expressed at higher levels in leaves and during seed germination than in roots and appeared to be not responsive to stress. The Aox expression profiles obtained from a M. truncatula microarray dataset also revealed a stress-induced co-expression of Aox1, Aox2b1 and Aox2b2 in leaves and roots. These results reinforce the stress-inducible co-expression of Aox1/Aox2b in some leguminous plants. Comparative genomic analysis indicates that this regulation is linked to Aox1/Aox2b proximity in the genome as a result of the gene rearrangement that occurred in some leguminous plants during evolution. The differential expression of Aox2b1/2b2 suggests that a second gene has been originated by recent gene duplication with neofunctionalization. © 2013 Elsevier GmbH. All rights reserved.

Note on the nonuniqueness of the massive Fierz-Pauli theory and spectator fields

It is possible to show that there are three independent families of models describing a massive spin-2 particle via a rank-2 tensor. One of them contains the massive Fierz-Pauli model, the only case described by a symmetric tensor. The three families have different local symmetries in the massless limit and can not be interconnected by any local field redefinition. We show here, however, that they can be related with the help of a decoupled and nondynamic (spectator) field. The spectator field may be either an antisymmetric tensor B μν=-Bνμ, a vector Aμ or a scalar field φ, corresponding to each of the three families. The addition of the extra field allows us to formulate master actions which interpolate between the symmetric Fierz-Pauli theory and the other models. We argue that massive gravity models based on the Fierz-Pauli theory are not expected to be equivalent to possible local self-interacting theories built up on top of the two new families of massive spin-2 models. The approach used here may be useful to investigate dual (nonsymmetric) formulations of higher-spin particles. © 2013 American Physical Society.

Evaluation of Estrogenic Potential of Flavonoids Using a Recombinant Yeast Strain and MCF7/BUS Cell Proliferation Assay

Phytoestrogens are of interest because of their reported beneficial effects on many human maladies including cancer, neurodegeneration, cardiovascular disease and diabetes. Furthermore, there is a search for compounds with estrogenic activity that can replace estrogen in hormone replacement therapy during menopause, without the undesirable effects of estrogen, such as the elevation of breast cancer occurrence. Thus, the principal objective of this study was to assess the estrogenic activity of flavonoids with different hydroxylation patterns: quercetin, kaempferol, luteolin, fisetin, chrysin, galangin, flavone, 3-hydroxyflavone, 5-hydroxyflavone and 7-hydroxyflavone via two different in vitro assays, the recombinant yeast assay (RYA) and the MCF-7 proliferation assay (E-screen), since the most potent phytoestrogens are members of the flavonoid family. In these assays, kaempferol was the only compound that showed ERα-dependent transcriptional activation activity by RYA, showing 6.74±1.7 nM EEQ, besides acting as a full agonist for the stimulation of proliferation of MCF-7/BUS cells. The other compounds did not show detectable levels of interaction with ER under the conditions used in the RYA. However, in the E-screen assay, compounds such as galangin, luteolin and fisetin also stimulated the proliferation of MCF-7/BUS cells, acting as partial agonists. In the evaluation of antiestrogenicity, the compounds quercetin, chrysin and 3-hydroxyflavone significantly inhibited the cell proliferation induced by 17-β-estradiol in the E-screen assay, indicating that these compounds may act as estrogen receptor antagonists. Overall, it became clear in the assay results that the estrogenic activity of flavonoids was affected by small structural differences such as the number of hydroxyl groups, especially those on the B ring of the flavonoid. © 2013 Resende et al.

Functions and vector fields on C(CPn)-singular manifolds

Let M2n+1 be a C(CPn) -singular manifold. We study functions and vector fields with isolated singularities on M2n+1. A C(CPn) -singular manifold is obtained from a smooth manifold M2n+1 with boundary in the form of a disjoint union of complex projective spaces CPn boolean OR CPn boolean OR ... boolean OR CPn with subsequent capture of a cone over each component of the boundary. Let M2n+1 be a compact C(CPn) -singular manifold with k singular points. The Euler characteristic of M2n+1 is equal to chi(M2n+1) = k(1 - n)/2. Let M2n+1 be a C(CPn)-singular manifold with singular points m(1), ..., m(k). Suppose that, on M2n+1, there exists an almost smooth vector field V (x) with finite number of zeros m(1), ..., m(k), x(1), ..., x(1). Then chi(M2n+1) = Sigma(l)(i=1) ind(x(i)) + Sigma(k)(i=1) ind(m(i)).

Geometrical hypercomplex coupling between electric and gravitational fields

The present work shows a coupling of electrical and gravitational fields through Cauchy-Riemann conditions for quaternions present in a previous paper [1]. It is also obtained an extended version of the Laplace-like equations for quaternions, now written in terms of both electric and gravitational fields.

Influence of occlusal contact area on cusp defection and stress distribution

The purpose of this study was to evaluate the effect of occlusal contact area for loading on the cuspal defection and stress distribution in a first premolar restored with a high elastic modulus restorative material. The Rhinoceros 4.0 software was used for modeling the three-dimensional geometries of dental and periodontal structures and the inlay restoration. Thus, two different models, intact and restored teeth with three occlusal contact areas, 0.1, 0.5 and 0.75 mm(2), on enamel at the occlusal surface of buccal and lingual cusps. Finite element analysis (FEA) was performed with the program ANSYS (Workbench 13.0), which generated a mesh with tetrahedral elements with greater refinement in the regions of interest, and was constrained at the bases of cortical and trabecular bone in all axis and loaded with 100 N normal to each contact area. To analysis of maximum principal stress, the smaller occlusal contact area showed greater compressive stress in region of load application for both the intact and inlay restored tooth. However, tensile stresses at the occlusal isthmus were similar for all three tested occlusal contact areas (60 MPa). To displacement of the cusps was higher for teeth with inlay (0.46-0.48 mm). For intact teeth, the smaller contact area showed greater displacement (0.10 mm). For teeth with inlays, the displacement of the cusps were similar in all types of occlusal area. Cuspal displacement was higher in the restored tooth when compared to the intact tooth, but there were no significant variations even with changes in the occlusal contact area. RELEVANCE CLINICAL: Occlusal contacts have a great influence on the positioning of teeth being able to maintain the position and stability of the mandible. Axial loads would be able to generate more uniform stress at the root presenting a greater concentration of load application in the point and the occlusal surface. Thus, is necessary to analyze the relationship between these occlusal contacts as dental wear and subsequent occlusal interferences.

Patients on the Waiting List for Liver Transplantation: Caregiver Burden and Stress

Over the last few decades, informal caregivers of patients with chronic diseases have received more attention, and there is a growing volume of studies demonstrating high rates of burden, stress, and mental disorders in this group of individuals. The objective of this study was to evaluate the burden, stress, and psychosocial characteristics of informal caregivers of liver transplantation candidates. Participants were assessed by individual evaluations with the following instruments: a semi-structured interview, the Caregiver Burden Scale, the Inventario de Sintomas de Stress para Adultos de Lipp, and the Beck Depression Inventory. The Mann-Whitney test was used for statistical analysis with a significance level of 0.05. The characteristics of the study group (n = 61) were similar to those of groups in other studies with respect to gender (82% were women), kinship (64% were spouses), and age (the mean age was 47.6 years). The main stressors identified by the participants were as follows: doubts about ways to react in a crisis or in emergency situations (42.6%), mood swings of the patient (29.5%), and care involving food and medications (27.9%). Approximately 25% of the caregivers reported that they felt unprepared to adequately perform their roles. Data analysis indicated a greater burden overall on caregivers when the patient`s Model for End-Stage Liver Disease score was greater than or equal to 15 points (P = 0.041). Furthermore, caregivers of patients with alcoholic liver disease showed higher depression (P = 0.034) and overall burden scores (P = 0.031) versus caregivers of patients with liver disease due to other etiologies. In conclusion, the participants showed significantly high levels of burden, stress, and depression. Support measures and caregiver preparation should be implemented by health care providers. Liver Transpl 16: 1164-1168, 2010. (C) 2010 AASLD.

Long-Term Occupational Exposure to Organic Solvents Affects Color Vision, Contrast Sensitivity and Visual Fields

The purpose of this study was to evaluate the visual outcome of chronic occupational exposure to a mixture of organic solvents by measuring color discrimination, achromatic contrast sensitivity and visual fields in a group of gas station workers. We tested 25 workers (20 males) and 25 controls with no history of chronic exposure to solvents (10 males). All participants had normal ophthalmologic exams. Subjects had worked in gas stations on an average of 9.6 +/- 6.2 years. Color vision was evaluated with the Lanthony D15d and Cambridge Colour Test (CCT). Visual field assessment consisted of white-on-white 24-2 automatic perimetry (Humphrey II-750i). Contrast sensitivity was measured for sinusoidal gratings of 0.2, 0.5, 1.0, 2.0, 5.0, 10.0 and 20.0 cycles per degree (cpd). Results from both groups were compared using the Mann-Whitney U test. The number of errors in the D15d was higher for workers relative to controls (p<0.01). Their CCT color discrimination thresholds were elevated compared to the control group along the protan, deutan and tritan confusion axes (p<0.01), and their ellipse area and ellipticity were higher (p<0.01). Genetic analysis of subjects with very elevated color discrimination thresholds excluded congenital causes for the visual losses. Automated perimetry thresholds showed elevation in the 9 degrees, 15 degrees and 21 degrees of eccentricity (p<0.01) and in MD and PSD indexes (p<0.01). Contrast sensitivity losses were found for all spatial frequencies measured (p<0.01) except for 0.5 cpd. Significant correlation was found between previous working years and deutan axis thresholds (rho = 0.59; p<0.05), indexes of the Lanthony D15d (rho = 0.52; p<0.05), perimetry results in the fovea (rho = -0.51; p<0.05) and at 3, 9 and 15 degrees of eccentricity (rho = -0.46; p<0.05). Extensive and diffuse visual changes were found, suggesting that specific occupational limits should be created.

Maternal factors regulating preterm infants' responses to pain and stress while in maternal kangaroo care

The relationship between maternal factors and the response of preterm infants to pain and stress experienced during heel puncture while in maternal kangaroo care was investigated. This descriptive study included 42 mothers and their preterm infants cared for in a neonatal unit. Data were collected in the baseline, procedure, and recovery phases. We measured the neonates' facial actions, sleep and wake states, crying, salivary cortisol levels, and heart rate, in addition to the mothers' behavior, salivary cortisol levels, and mental condition. The influence of the maternal explanatory variables on the neonatal response variables were verified through bivariate analysis, ANOVA, and multiple regression. The mothers' behavior and depression and/or anxiety did not affect the neonates' responses to pain and stress, though the mothers' levels of salivary cortisol before the procedure explained the variance in the neonates' levels of salivary cortisol after the procedure (p=0.036). Additionally, the mothers' baseline levels of salivary cortisol along with the neonates' age explained the variance in the neonates' heart rate (p=0.001). The ability of mothers to regulate their own stress contributed to the infants' responses to pain and stress.

Effects of cryogenic and stress relief treatments on temper carbide precipitation in AISI D2 tool steel

The effects of cryogenic and stress relief treatments on temper carbide precipitation in the cold work tool steel AISI D2 were studied. For the cryogenic treatment the temperature was −196°C and the holding time was 2, 24 or 30 h. The stress relief heat treatment was carried at 130°C/90 min, when applied. All specimens were compared to a standard thermal cycle. Specimens were studied using metallographic characterisation, X-ray diffraction and thermoelectric power measurements. The metallographic characterisation used SEM (scanning electron microscopy) and SEM-FEG (SEM with field emission gun), besides OM (optical microscopy). No variation in the secondary carbides (micrometre sized) precipitation was found. The temper secondary carbides (nanosized) were found to be more finely dispersed in the matrix of the specimens with cryogenic treatment and without stress relief. The refinement of the temper secondary carbides was attributed to a possible in situ carbide precipitation during tempering.

Jaw mechanics of the pterosaur skull construction and the evolution of toothlessness

In the present study, pterosaur skull constructions were analysed using a combined approach of finite element analysis (FEA), static investigations as well as applying classical beam theory and lever mechanics. The study concentrates on the operating regime „bite“, where loads are distributed via the dentition or a keratinous rhamphotheca into the skull during jaw occlusion. As a first step, pterosaur tooth constructions were analysed. The different morphologies of the tooth construction determine specific operational ranges, in which the teeth perform best (= greatest resistance against failure). The incomplete enamel-covering of the pterosaur tooth constructions thereby leads to a reduction of strain and stress and to a greater lateral elasticity than for a complete enamel cover. This permits the development of high and lateral compressed tooth constructions. Further stress-absorption occurs in the periodontal membrane, although its mechanical properties can not be clarified unambiguously. A three-dimensionally preserved skull of Anhanguera was chosen as a case-study for the investigation of the skull constructions. CT-scans were made to get information about the internal architecture, supplemented by thin-sections of a rostrum of a second Anhanguera specimen. These showed that the rostrum can be approximated as a double-walled triangular tube with a large central vacuity and an average wall-thickness of the bony layers of about 1 mm. On base of the CT-scans, a stereolithography of the skull of Anhanguera was made on which the jaw adductor and abductor muscles were modelled, permitting to determine muscular forces. The values were used for the lever mechanics, cantilever and space frame analysis. These studies and the FEA show, that the jaw reaction forces are critical for the stability of the skull construction. The large jugal area ventral to the orbita and the inclined occipital region act as buttresses against these loads. In contrast to the orbitotemporal region which is subject to varying loading conditions, the pattern in the rostrum is less complex. Here, mainly bending in dorsal direction and torsion occur. The hollow rostrum leads to a reduction of weight of the skull and to a high bending and torsional resistance. Similar to the Anhanguera skull construction, the skulls of those pterosaur taxa were analysed, from which enough skull material is know to permit a reliable reconstruction. Furthermore, FEA were made from five selected taxa. The comparison of the biomechanical behaviour of the different skull constructions results in major transformational processes: elongation of rostra, inclination of the occipital region, variation of tooth morphology, reduction of the dentition and replacement of teeth by a keratinous hook or rhamphotheca, fusion of naris and antorbital fenestra, and the development of bony and soft-tissue crests. These processes are discussed for their biomechanical effects during bite. Certain optional operational ranges for feeding are assigned to the different skull constructions and previous hypotheses (e.g. skimming) are verified. Using the principle of economisation, these processes help to establish irreversible transformations and to define possible evolutionary pathways. The resulting constructional levels and the structural variations within these levels are interpreted in light of a greater feeding efficiency and reduction of bony mass combined with an increased stability against the various loads. The biomechanical conclusive pathways are used for comparison and verification of recent hypothesis of the phylogenetic systematics of pterosaurs.

Fault delineation and stress orientations from the analysis of background, low magnitude seismicity in Southern Apennines (Italy)

The aim of this work was to show that refined analyses of background, low magnitude seismicity allow to delineate the main active faults and to accurately estimate the directions of the regional tectonic stress that characterize the Southern Apennines (Italy), a structurally complex area with high seismic potential. Thanks the presence in the area of an integrated dense and wide dynamic network, was possible to analyzed an high quality microearthquake data-set consisting of 1312 events that occurred from August 2005 to April 2011 by integrating the data recorded at 42 seismic stations of various networks. The refined seismicity location and focal mechanisms well delineate a system of NW-SE striking normal faults along the Apenninic chain and an approximately E-W oriented, strike-slip fault, transversely cutting the belt. The seismicity along the chain does not occur on a single fault but in a volume, delimited by the faults activated during the 1980 Irpinia M 6.9 earthquake, on sub-parallel predominant normal faults. Results show that the recent low magnitude earthquakes belongs to the background seismicity and they are likely generated along the major fault segments activated during the most recent earthquakes, suggesting that they are still active today thirty years after the mainshock occurrences. In this sense, this study gives a new perspective to the application of the high quality records of low magnitude background seismicity for the identification and characterization of active fault systems. The analysis of the stress tensor inversion provides two equivalent models to explain the microearthquake generation along both the NW-SE striking normal faults and the E- W oriented fault with a dominant dextral strike-slip motion, but having different geological interpretations. We suggest that the NW-SE-striking Africa-Eurasia convergence acts in the background of all these structures, playing a primary and unifying role in the seismotectonics of the whole region.

A Design Method for Flexure-Based Compliant Mechanisms on the Basis of Stiffness and Stress Characteristics

Geometric nonlinearities of flexure hinges introduced by large deflections often complicate the analysis of compliant mechanisms containing such members, and therefore, Pseudo-Rigid-Body Models (PRBMs) have been well proposed and developed by Howell [1994] to analyze the characteristics of slender beams under large deflection. These models, however, fail to approximate the characteristics for the deep beams (short beams) or the other flexure hinges. Lobontiu's work [2001] contributed to the diverse flexure hinge analysis building on the assumptions of small deflection, which also limits the application range of these flexure hinges and cannot analyze the stiffness and stress characteristics of these flexure hinges for large deflection. Therefore, the objective of this thesis is to analyze flexure hinges considering both the effects of large-deflection and shear force, which guides the design of flexure-based compliant mechanisms. The main work conducted in the thesis is outlined as follows. 1. Three popular types of flexure hinges: (circular flexure hinges, elliptical flexure hinges and corner-filleted flexure hinges) are chosen for analysis at first. 2. Commercial software (Comsol) based Finite Element Analysis (FEA) method is then used for correcting the errors produced by the equations proposed by Lobontiu when the chosen flexure hinges suffer from large deformation. 3. Three sets of generic design equations for the three types of flexure hinges are further proposed on the basis of stiffness and stress characteristics from the FEA results. 4. A flexure-based four-bar compliant mechanism is finally studied and modeled using the proposed generic design equations. The load-displacement relationships are verified by a numerical example. The results show that a maximum error about the relationship between moment and rotation deformation is less than 3.4% for a flexure hinge, and it is lower than 5% for the four-bar compliant mechanism compared with the FEA results.

Using national FADN database to describe Italian farms and arable fields. A comparison of sustainability level between organic versus conventional regimes.

The study defines a new farm classification and identifies the arable land management. These aspects and several indicators are taken into account to estimate the sustainability level of farms, for organic and conventional regimes. The data source is Italian Farm Account Data Network (RICA) for years 2007-2011, which samples structural and economical information. An environmental data has been added to the previous one to better describe the farm context. The new farm classification describes holding by general informations and farm structure. The general information are: adopted regime and farm location in terms of administrative region, slope and phyto-climatic zone. The farm structures describe the presence of main productive processes and land covers, which are recorded by FADN database. The farms, grouped by homogeneous farm structure or farm typology, are evaluated in terms of sustainability. The farm model MAD has been used to estimate a list of indicators. They describe especially environmental and economical areas of sustainability. Finally arable lands are taken into account to identify arable land managements and crop rotations. Each arable land has been classified by crop pattern. Then crop rotation management has been analysed by spatial and temporal approaches. The analysis reports a high variability inside regimes. The farm structure influences indicators level more than regimes, and it is not always possible to compare the two regimes. However some differences between organic and conventional agriculture have been found. Organic farm structures report different frequency and geographical location than conventional ones. Also different connections among arable lands and farm structures have been identified.

Atmospheric plasma processes for microbial inactivation: food applications and stress response in Listeria monocytogenes

This PhD thesis is focused on cold atmospheric plasma treatments (GP) for microbial inactivation in food applications. In fact GP represents a promising emerging technology alternative to the traditional methods for the decontamination of foods. The objectives of this work were to evaluate: - the effects of GP treatments on microbial inactivation in model systems and in real foods; - the stress response in L. monocytogenes following exposure to different GP treatments. As far as the first aspect, inactivation curves were obtained for some target pathogens, i.e. Listeria monocytogenes and Escherichia coli, by exposing microbial cells to GP generated with two different DBD equipments and processing conditions (exposure time, material of the electrodes). Concerning food applications, the effects of different GP treatments on the inactivation of natural microflora and Listeria monocytogenes, Salmonella Enteritidis and Escherichia coli on the surface of Fuji apples, soya sprouts and black pepper were evaluated. In particular the efficacy of the exposure to gas plasma was assessed immediately after treatments and during storage. Moreover, also possible changes in quality parameters such as colour, pH, Aw, moisture content, oxidation, polyphenol-oxidase activity, antioxidant activity were investigated. Since the lack of knowledge of cell targets of GP may limit its application, the possible mechanism of action of GP was studied against 2 strains of Listeria monocytogenes by evaluating modifications in the fatty acids of the cytoplasmic membrane (through GC/MS analysis) and metabolites detected by SPME-GC/MS and 1H-NMR analyses. Moreover, changes induced by different treatments on the expression of selected genes related to general stress response, virulence or to the metabolism were detected with Reverse Transcription-qPCR. In collaboration with the Scripps Research Institute (La Jolla, CA, USA) also proteomic profiles following gas plasma exposure were analysed through Multidimensional Protein Identification Technology (MudPIT) to evaluate possible changes in metabolic processes.