Defects in vesicle core induced by Escherichia coli dihydroorotate dehydrogenase

Dihydroorotate dehydrogenase (DHODH) catalyzes the oxidation of dihydroorotate to orotate during the fourth step of the de novo pyrimidine synthesis pathway. In rapidly proliferating mammalian cells, pyrimidine salvage pathway is insufficient to overcome deficiencies in that pathway for nucleotide synthesis. Moreover, as certain parasites lack salvage enzymes, relying solely on the de novo pathway, DHODH inhibition has turned out as an efficient way to block pyrimidine biosynthesis. Escherichia coli DHODH (EcDHODH) is a class 2 DHODH, found associated to cytosolic membranes through an N-terminal extension. We used electronic spin resonance (ESR) to study the interaction of EcDHODH with vesicles of 1,2-dioleoyl-sn-glycero-phosphatidylcholine/detergent. Changes in vesicle dynamic structure induced by the enzyme were monitored via spin labels located at different positions of phospholipid derivatives. Two-component ESR spectra are obtained for labels 5- and 1 0-phosphatidylcholine in presence of EcDHODH, whereas other probes show a single-component spectrum. The appearance of an additional spectral component with features related to fast-motion regime of the probe is attributed to the formation of a defect-like structure in the membrane hydrophobic region. This is probably the mechanism used by the protein to capture quinones used as electron acceptors during catalysis. The use of specific spectral simulation routines allows us to characterize the ESR spectra in terms of changes in polarity and mobility around the spin-labeled phospholipids. We believe this is the first report of direct evidences concerning the binding of class 2 DHODH to membrane systems.

Characterization of Yeast Extracellular Vesicles: Evidence for the Participation of Different Pathways of Cellular Traffic in Vesicle Biogenesis

Background: Extracellular vesicles in yeast cells are involved in the molecular traffic across the cell wall. In yeast pathogens, these vesicles have been implicated in the transport of proteins, lipids, polysaccharide and pigments to the extracellular space. Cellular pathways required for the biogenesis of yeast extracellular vesicles are largely unknown. Methodology/Principal Findings: We characterized extracellular vesicle production in wild type (WT) and mutant strains of the model yeast Saccharomyces cerevisiae using transmission electron microscopy in combination with light scattering analysis, lipid extraction and proteomics. WT cells and mutants with defective expression of Sec4p, a secretory vesicle-associated Rab GTPase essential for Golgi-derived exocytosis, or Snf7p, which is involved in multivesicular body (MVB) formation, were analyzed in parallel. Bilayered vesicles with diameters at the 100-300 nm range were found in extracellular fractions from yeast cultures. Proteomic analysis of vesicular fractions from the cells aforementioned and additional mutants with defects in conventional secretion pathways (sec1-1, fusion of Golgi-derived exocytic vesicles with the plasma membrane; bos1-1, vesicle targeting to the Golgi complex) or MVB functionality (vps23, late endosomal trafficking) revealed a complex and interrelated protein collection. Semi-quantitative analysis of protein abundance revealed that mutations in both MVB- and Golgi-derived pathways affected the composition of yeast extracellular vesicles, but none abrogated vesicle production. Lipid analysis revealed that mutants with defects in Golgi-related components of the secretory pathway had slower vesicle release kinetics, as inferred from intracellular accumulation of sterols and reduced detection of these lipids in vesicle fractions in comparison with WT cells. Conclusions/Significance: Our results suggest that both conventional and unconventional pathways of secretion are required for biogenesis of extracellular vesicles, which demonstrate the complexity of this process in the biology of yeast cells.

Proteoliposomes Harboring Alkaline Phosphatase and Nucleotide Pyrophosphatase as Matrix Vesicle Biomimetics

We have established a proteoliposome system as an osteoblast-derived matrix vesicle (MV) biomimetic to facilitate the study of the interplay of tissue-nonspecific alkaline phosphatase (TNAP) and NPP1 (nucleotide pyrophosphatase/phosphodiesterase-1) during catalysis of biomineralization substrates. First, we studied the incorporation of TNAP into liposomes of various lipid compositions (i.e. in pure dipalmitoyl phosphatidylcholine (DPPC), DPPC/dipalmitoyl phosphatidylserine (9:1 and 8:2), and DPPC/dioctadecyl-dimethylammonium bromide (9:1 and 8:2) mixtures. TNAP reconstitution proved virtually complete in DPPC liposomes. Next, proteoliposomes containing either recombinant TNAP, recombinant NPP1, or both together were reconstituted in DPPC, and the hydrolysis of ATP, ADP, AMP, pyridoxal-5`-phosphate (PLP), p-nitrophenyl phosphate, p-nitrophenylthymidine 5`-monophosphate, and PP(i) by these proteoliposomes was studied at physiological pH. p-Nitrophenylthymidine 5`-monophosphate and PLP were exclusively hydrolyzed by NPP1-containing and TNAP-containing proteoliposomes, respectively. In contrast, ATP, ADP, AMP, PLP, p-nitrophenyl phosphate, and PPi were hydrolyzed by TNAP-, NPP1-, and TNAP plus NPP1- containing proteoliposomes. NPP1 plus TNAP additively hydrolyzed ATP, but TNAP appeared more active in AMP formation than NPP1. Hydrolysis of PPi by TNAP-, and TNAP plus NPP1- containing proteoliposomes occurred with catalytic efficiencies and mild cooperativity, effects comparable with those manifested by murine osteoblast-derived MVs. The reconstitution of TNAP and NPP1 into proteoliposome membranes generates a phospholipid microenvironment that allows the kinetic study of phosphosubstrate catabolism in a manner that recapitulates the native MV microenvironment.

Flavoring agents present in a dentifrice can modify volatile sulphur compounds (VSCs) formation in morning bad breath

This study aimed to evaluate the effects of a flavor-containing dentifrice on the formation of volatile sulphur compounds (VSCs) in morning bad breath. A two-step, blinded, crossover, randomized study was carried out in 50 dental students with a healthy periodontium divided into two experimental groups: flavor-containing dentifrice (test) and non-flavor-containing dentifrice (control). The volunteers received the designated dentifrice and a new toothbrush for a 3 X/day brushing regimen for 2 periods of 30 days. A seven-day washout interval was used between the periods. The assessed parameters were: plaque index (PI), gingival index (GI), organoleptic breath scores (ORG), VSC levels (as measured by a portable sulphide monitor) before (H1) and after (H2) cleaning of the tongue, tongue coating (TC) wet weight and BANA test from TC samples. The intra-group analysis showed a decrease in ORG, from 3 to 2, after 30 days for the test group (p < 0.05). The inter-group analysis showed lower values in ORG, H1 and H2 for the test group (p < 0.05). There was no difference between the amount of TC between groups and the presence of flavor also did not interfere in the BANA results between groups (p > 0.05). These findings suggest that a flavor-containing dentifrice seems to prevent VSCs formation in morning bad breath regardless of the amount of TC in periodontally healthy subjects.

Endodontic treatment of dental formation anomalies

PURPOSE: Dental fusion is defined as the union of two dental germs at some stage of their development. The aim of this article is to report the endodontic treatment of two clinical cases of dental fusion. CASE DESCRIPTION: In the first case, the patient was referred by an orthodontist for endodontic treatment of tooth 12, which was fused to 13. Surgical separation and later replacement of the involved elements in the dental arch was indicated. In the second case, the patient sought dental attendance due to spontaneous pain. In the radiographic exam, gemination in tooth 11 and fusion of 21 with a supernumerary tooth was observed. The fused teeth were endodontically treated, and patients were referred to other dental specialties to reestablish esthetics and function. CONCLUSION: The dentist must be able to diagnose, differentiate and treat these dental anomalies adequately, with the goal of maintaining patients' oral health.

Characterization of oil shale residue and rejects from irati formation by electron paramagnetic resonance

In this study, sedimentary organic matter of oil shale rejects, calschist, shale fine and the so called retorted shale from Irati formation was characterized. EPR was used to analyse the samples regarding loss of signal in g = 2.003 associated to the organic free radical with the calcined samples and washing with hydrogen peroxide. The radical signal was detected in all samples, however, for the calschist and shale fine samples another signal was identified at g = 2.000 which disappeared when the sample was heated at 400 ºC. Hydrogen peroxide washing was also performed and it was noted that after washing the signal appeared around g = 2.000 for all samples, including retorted shale, which might be due to the quartz E1 defect.

Marine influence in the Barreiras Formation, State of Alagoas, northeastern Brazil

Tidal processes were important for deposition of the Barreiras Formation located in northern Brazil, while correlatable deposits in northeastern Brazil have been traditionally related to continental environments. Facies analysis in southern Alagoas revealed that the Barreiras Formation consists of cross-stratified conglomerates and sandstones (facies Sx and Cgx), compound cross-stratified sandstones (facies Cx), and heterolithic beddings (facies H). A significant portion of these deposits occurs within channel morphologies displaying fining and thinning upward successions. An abundance of sedimentary features is comparable to those from the northern Brazilian counterpart. These include: tidal bundles; herringbone cross-stratification; heterolithic beddings with sandstone and mudstone beds in sharp contacts; and ichnofossils mostly consisting of Ophiomorpha nodosa, Skolithos and Planolites. Altogether, these features point to a marginal marine depositional setting dominated by tidal processes, which are related to an estuarine system, an interpretation also provided for the Barreiras Formation in northern Brazil. The widespread occurrence of deposits with unambiguous evidence of tidal processes in the Barreiras Formation of northern Brazil, and now in the State of Alagoas, leads to argue that the early/middle Miocene worldwide marine transgression might have left a much more widespread sedimentary record along the Brazilian coast than currently regarded.

Os histricomorfos sul-americanos: uma análise comparativa do desenvolvimento embriológico

O objetivo do estudo foi realizar uma análise embriológica dos roedores histricomorfos (paca, cutia, preá e capivara), a fim de comparar com a de outros roedores e com a morfogênese humana um padrão embriológico. Utilizaram-se 8 espécimes de roedores sendo, 2 embriões para cada espécie coletada, ambas em inicio de gestação. Estes foram retirados dos úteros gestantes através de ovariosalpingohisterectomia parcial, seguido de fixação em solução de paraformaldeído 4%. Para as mensurações de Crow-Rump, adotou-se como referência a crista nucal numa extremidade e da última vértebra sacral na extremidade oposta. De forma geral, os embriões analisados mostraram as seguintes características morfológicas: divisão dos arcos branquiais, o não fechamento do neuróporo cranial em alguns embriões estudados, a curvatura cranial acentuada e os somitos delimitados e individualizados. O broto dos membros apresentava-se em desenvolvimento em formato de remo, além da impressão cardíaca e fígado. Na região caudal, visualizou-se a curvatura crânio-caudal, a vesícula óptica com e sem pigmentação da retina, a abertura do tubo neural na região do quarto ventrículo encefálico, a fosseta nasal e a formação das vesículas encefálicas. Concluímos que desenvolvimento embriológico dos roedores histricomorfos pode ser comparado à morfogênese de ratos, cobaios, coelhos e humanos nos diferentes estágios de desenvolvimento, tomando apenas o cuidado com as particularidades de cada espécie, além da implementação de tecnologias reprodutivas, especialmente a de embriões, a qual requer o conhecimento do desenvolvimento pré-implantação referente às fases de desenvolvimento.

Ecdysteroid-Dependent Expression of the Tweedle and Peroxidase Genes during Adult Cuticle Formation in the Honey Bee, Apis mellifera

Cuticle renewal is a complex biological process that depends on the cross talk between hormone levels and gene expression. This study characterized the expression of two genes encoding cuticle proteins sharing the four conserved amino acid blocks of the Tweedle family, AmelTwdl1 and AmelTwdl2, and a gene encoding a cuticle peroxidase containing the Animal haem peroxidase domain, Ampxd, in the honey bee. Gene sequencing and annotation validated the formerly predicted tweedle genes, and revealed a novel gene, Ampxd, in the honey bee genome. Expression of these genes was studied in the context of the ecdysteroid-coordinated pupal-to-adult molt, and in different tissues. Higher transcript levels were detected in the integument after the ecdysteroid peak that induces apolysis, coinciding with the synthesis and deposition of the adult exoskeleton and its early differentiation. The effect of this hormone was confirmed in vivo by tying a ligature between the thorax and abdomen of early pupae to prevent the abdominal integument from coming in contact with ecdysteroids released from the prothoracic gland. This procedure impaired the natural increase in transcript levels in the abdominal integument. Both tweedle genes were expressed at higher levels in the empty gut than in the thoracic integument and trachea of pharate adults. In contrast, Ampxd transcripts were found in higher levels in the thoracic integument and trachea than in the gut. Together, the data strongly suggest that these three genes play roles in ecdysteroid-dependent exoskeleton construction and differentiation and also point to a possible role for the two tweedle genes in the formation of the cuticle (peritrophic membrane) that internally lines the gut.

Effects of Low-Level Laser Therapy on Pain and Scar Formation After Inguinal Herniation Surgery: A Randomized Controlled Single-Blind Study

Objective: The aim of this study was to investigate the efficacy of an infrared GaAlAs laser operating with a wavelength of 830 nm in the postsurgical scarring process after inguinal-hernia surgery. Background: Low-level laser therapy (LLLT) has been shown to be beneficial in the tissue-repair process, as previously demonstrated in tissue culture and animal experiments. However, there is lack of studies on the effects of LLLT on postsurgical scarring of incisions in humans using an infrared 830-nm GaAlAs laser. Method: Twenty-eight patients who underwent surgery for inguinal hernias were randomly divided into an experimental group (G1) and a control group (G2). G1 received LLLT, with the first application performed 24 h after surgery and then on days 3, 5, and 7. The incisions were irradiated with an 830-nm diode laser operating with a continuous power output of 40 mW, a spot-size aperture of 0.08 cm(2) for 26 s, energy per point of 1.04 J, and an energy density of 13 J/cm(2). Ten points per scar were irradiated. Six months after surgery, both groups were reevaluated using the Vancouver Scar Scale (VSS), the Visual Analog Scale, and measurement of the scar thickness. Results: G1 showed significantly better results in the VSS totals (2.14 +/- 1.51) compared with G2 (4.85 +/- 1.87); in the thickness measurements (0.11 cm) compared with G2 (0.19 cm); and in the malleability (0.14) compared with G2 (1.07). The pain score was also around 50% higher in G2. Conclusion: Infra-red LLLT (830 nm) applied after inguinal-hernia surgery was effective in preventing the formation of keloids. In addition, LLLT resulted in better scar appearance and quality 6 mo postsurgery.

Star formation efficiency in galaxy clusters

Context. The luminous material in clusters of galaxies exists in two forms: the visible galaxies and the X-ray emitting intra-cluster medium. The hot intra-cluster gas is the major observed baryonic component of clusters, about six times more massive than the stellar component. The mass contained within visible galaxies is approximately 3% of the dynamical mass. Aims. Our aim was to analyze both baryonic components, combining X-ray and optical data of a sample of five galaxy clusters (Abell 496, 1689, 2050, 2631 and 2667), within the redshift range 0.03 < z < 0.3. We determined the contribution of stars in galaxies and the intra-cluster medium to the total baryon budget. Methods. We used public XMM-Newton data to determine the gas mass and to obtain the X-ray substructures. Using the optical counterparts from SDSS or CFHT we determined the stellar contribution. Results. We examine the relative contribution of galaxies, intra-cluster light and intra-cluster medium to baryon budget in clusters through the stellar-to-gas mass ratio, estimated with recent data. We find that the stellar-to-gas mass ratio within r(500) (the radius within which the mean cluster density exceeds the critical density by a factor of 500), is anti-correlated with the ICM temperature, which range from 24% to 6% while the temperature ranges from 4.0 to 8.3 keV. This indicates that less massive cold clusters are more prolific star forming environments than massive hot clusters.

High spatial resolution spectroscopy of W51IRS 2E and IRS 2W: Two very massive young stars in early formation stages

We present K-band spectra of the near infrared counterparts to IRS 2E and IRS 2W which is associated with the ultracompact H II region W51d, both of them embedded sources in the Galactic compact H II region W51 IRS 2. The high spatial resolution observations were obtained with the laser guide star facility and Near-infrared Integral Field Spectrograph (NIFS) mounted at the Gemini-North observatory. The spectrum of the ionizing source of W51d shows the photospheric features N III ( 21155 angstrom) in emission and He II ( 21897 angstrom) in absorption which lead us to classify it as a young O3 type star. We detected CO overtone in emission at 23000 angstrom in the spectrum of IRS 2E, suggesting that it is a massive young object still surrounded by an accretion disk, probably transitioning from the hot core phase to an ultracompact H II region.

Searching for star formation outside galaxies: Multiwavelength analysis of the intragroup medium of Hickson Compact Group 100

We use multiwavelength data (H I, FUV, NUV, R) to search for evidence of star formation in the intragroup medium of the Hickson Compact Group 100. We find that young star-forming regions are located in the intergalactic H I clouds of the compact group which extend to over 130 kpc away from the main galaxies. A tidal dwarf galaxy (TDG) candidate is located in the densest region of the H I tail, 61 kpc from the brightest group member and its age is estimated to be only 3.3 Myr. Fifteen other intragroup H II regions and TDG candidates are detected in the Galaxy Evolution Explorer (GALEX) FUV image and within a field 10' x 10' encompassing the H I tail. They have ages <200 Myr, H I masses of 10(9.2-10.4) M(circle dot), 0.001

Two formation channels of ultra-compact dwarf galaxies in Hickson compact groups

Context. The formation of ultra-compact dwarf galaxies (UCDs) is believed to be driven by interaction, and UCDs are abundant in the cores of galaxy clusters, environments that mark the end-point of galaxy evolution. Nothing is known about the properties of UCDs in compact groups of galaxies, environments where most of galaxy evolution and interaction is believed to occur and where UCDs in an intermediate stage in their evolution may be expected. Aims. The main goal of this study is to detect and characterize, for the first time, the UCD population of compact groups of galaxies. For that, two nearby groups in different evolutionary stages, HCG22 and HCG90, were targeted. Methods. We selected about 40 UCD candidates from pre-existing photometry of both groups, and obtained spectra of these candidates using the VLT FORS2 instrument in MXU mode. Archival HST/ACS imaging was used to measure their structural parameters. Results. We detect 16 and 5 objects belonging to HCG22 and HCG90, respectively, covering the magnitude range -10.0 > M(R) > -11.5 mag. Their integrated colours are consistent with old ages covering a broad range in metallicities (metallicities confirmed by the spectroscopic measurements). Photometric mass estimates put 4 objects in HCG90 and 9 in HCG22 in the mass range of UCDs (> 2 x 10(6) M(circle dot)) for an assumed age of 12Gyr. These UCDs are on average 2-3 times larger than the typical size of Galactic GCs, covering a range of 2 less than or similar to r(h) less than or similar to 21 pc. The UCDs in HCG22 are more concentrated around the central galaxy than in HCG90, at the 99% confidence level. They cover a broad range in [alpha/Fe] abundances from sub-to super-solar. The spectra of 3 UCDs (2 in HCG22, 1 in HCG90) show tentative evidence of intermediate age stellar populations. The clearest example is the largest and most massive UCD (similar to 10(7) M(circle dot)) in our sample, which is detected in HCG22. Its properties are most consistent with a stripped dwarf galaxy nucleus. We calculate the specific frequency (S(N)) of UCDs for both groups, finding that HCG22 has about three times higher S(N) than HCG90. Conclusions. The ensemble properties of the detected UCDs supports two co-existing formation channels: a star cluster origin (low-luminosity, compact sizes, old ages, super-solar alpha/Fe), and an origin as tidally stripped dwarf nuclei (more extended and younger stellar populations). Our results imply that the UCDs detected in both groups do not, in their majority, originate from relatively recent galaxy interactions. Most of the detected UCDs have likely been brought into the group along with their host galaxies.

Insights on the Milky Way bulge formation from the correlations between kinematics and metallicity

Context. Two main scenarios for the formation of the Galactic bulge are invoked, the first one through gravitational collapse or hierarchical merging of subclumps, the second through secular evolution of the Galactic disc. Aims. We aim to constrain the formation of the Galactic bulge through studies of the correlation between kinematics and metallicities in Baade's Window (l = 1 degrees, b = -4 degrees) and two other fields along the bulge minor axis (l = 0 degrees, b = -6 degrees and b = -12 degrees). Methods. We combine the radial velocity and the [Fe/H] measurements obtained with FLAMES/GIRAFFE at the VLT with a spectral resolution of R = 20 000, plus for the Baade's Window field the OGLE-II proper motions, and compare these with published N-body simulations of the Galactic bulge. Results. We confirm the presence of two distinct populations in Baade's Window found in Hill et al. (2010, A&A, submitted): the metal-rich population presents bar-like kinematics while the metal-poor population shows kinematics corresponding to an old spheroid or a thick disc. In this context the metallicity gradient along the bulge minor axis observed by Zoccali et al. (2008, A&A, 486, 177), visible also in the kinematics, can be related to a varying mix of these two populations as one moves away from the Galactic plane, alleviating the apparent contradiction between the kinematic evidence of a bar and the existence of a metallicity gradient. Conclusions. We show evidence that the two main scenarios for the bulge formation co-exist within the Milky Way bulge.