Glut1 and Glut3 as Potential Prognostic Markers for Oral Squamous Cell Carcinoma

We associated clinical-pathological features of 142 OSCC with the expression pattern of GLUT1 and GLUT3 in order to estimate their prognostic value. Methods: Clinical-pathological features and overall survival data of 142 patients with Oral Squamous Cell Carcinoma (OSCC) were retrospectively reviewed from A. C. Camargo hospital records. A tissue microarray (TMA) was built for the immunohistochemical (IHC) analysis of GLUT 1 and GLUT 3. IHC results were evaluated according to the staining pattern and number of positive cells. Results: GLUT 1 was over expressed in 50.3% of OSSC cases showing membrane staining pattern. However, nuclear expression was observed in 49.7% of the analyzed cases. GLUT 3 over expression was detected in 21.1% of OSCC cases. The pattern of GLUT 1 expression showed significant association with alcohol consumption (p = 0.004). Positive cell membrane GLUT 3 protein expression was associated with advanced clinic-staging of tumours (p = 0.005) as well as with vascular embolization (p = 0.005). Positive expression of GLUT 3 was associated with unfavorable free-disease survival (p = 0.021). Conclusion: GLUT1 and GLUT3 protein expression evaluated by immunohistochemistry are, significantly, indicators of poor prognosis outcome in oral squamous cell carcinoma, probably due to the enhanced glycolytic metabolism of more aggressive neoplastic cells.

New Source of Muscle-Derived Stem Cells with Potential for Alveolar Bone Reconstruction in Cleft Lip and/or Palate Patients

Cleft lip and palate (CLP), one of the most frequent congenital malformations, affects the alveolar bone in the great majority of the cases, and the reconstruction of this defect still represents a challenge in the rehabilitation of these patients. One of the current most promising strategy to achieve this goal is the use of bone marrow stem cells (BMSC); however, isolation of BMSC or iliac bone, which is still the mostly used graft in the surgical repair of these patients, confers site morbidity to the donor. Therefore, in order to identify a new alternative source of stem cells with osteogenic potential without conferring morbidity to the donor, we have used orbicular oris muscle (OOM) fragments, which are regularly discarded during surgery repair (cheiloplasty) of CLP patients. We obtained cells from OOM fragments of four unrelated CLP patients (CLPMDSC) using previously described preplating technique. These cells, through flow cytometry analysis, were mainly positively marked for five mesenchymal stem cell antigens (CD29, CD90, CD105, SH3, and SH4), while negative for hematopoietic cell markers, CD14, CD34, CD45, and CD117, and for endothelial cell marker, CD31. After induction under appropriate cell culture conditions, these cells were capable to undergo chondrogenic, adipogenic, osteogenic, and skeletal muscle cell differentiation, as evidenced by immunohistochemistry. We also demonstrated that these cells together with a collagen membrane lead to bone tissue reconstruction in a critical-size cranial defects previously induced in non-immunocompromised rats. The presence of human DNA in the new bone was confirmed by PCR with human-specific primers and immunohistochemistry with human nuclei antibodies. In conclusion, we showed that cells from OOM have phenotypic and behavior characteristics similar to other adult stem cells, both in vitro and in vivo. Our findings suggest that these cells represent a promising source of stem cells for alveolar bone grafting treatment, particularly in young CLP patients.

Developmental Potential of Bovine Hand-Made Clone Embryos Reconstructed by Aggregation or Fusion with Distinct Cytoplasmic Volumes

Animal cloning has been associated with developmental abnormalities, with the level of heteroplasmy caused by the procedure being one of its potential limiting factors. The aim of this study was to determine the effect of the fusion of hemicytoplasts or aggregation of hemiembryos, varying the final cytoplasmic volume, on development and cell density of embryos produced by hand-made cloning (HMC), parthenogenesis or by in vitro fertilization (IVF). One or two enucleated hemicytoplasts were paired and fused with one skin somatic cell. Activated clone and zona-free parthenote embryos and hemiembryos were in vitro cultured in the well-of-the-well (WOW) system, being allocated to one of six experimental groups, on a per WOW basis: single clone or parthenote hemiembryos (1 x 50%); aggregation of two (2 x 50%), three (3 x 50%), or four (4 x 50%) clone or parthenote hemiembryos; single clone or parthenote embryos (1 x 100%); or aggregation of two clone or parthenote embryos (2 x 100%). Control zona-intact parthenote or IVF embryos were in vitro cultured in four-well dishes. Results indicated that the increase in the number of aggregated structures within each WOW was followed by a linear increase in cleavage, blastocyst rate, and cell density. The increase in cytoplasmic volume, either by fusion or by aggregation, had a positive effect on embryo development, supporting the establishment of pregnancies and the birth of a viable clone calf after transfer to recipients. However, embryo aggregation did not improve development on a hemicytoplast basis, except for the aggregation of two clone embryos.

Sequential analysis of global gene expression profiles in immature and in vitro matured bovine oocytes: potential molecular markers of oocyte maturation

Background: Without intensive selection, the majority of bovine oocytes submitted to in vitro embryo production (IVP) fail to develop to the blastocyst stage. This is attributed partly to their maturation status and competences. Using the Affymetrix GeneChip Bovine Genome Array, global mRNA expression analysis of immature (GV) and in vitro matured (IVM) bovine oocytes was carried out to characterize the transcriptome of bovine oocytes and then use a variety of approaches to determine whether the observed transcriptional changes during IVM was real or an artifact of the techniques used during analysis. Results: 8489 transcripts were detected across the two oocyte groups, of which similar to 25.0% (2117 transcripts) were differentially expressed (p < 0.001); corresponding to 589 over-expressed and 1528 under-expressed transcripts in the IVM oocytes compared to their immature counterparts. Over expression of transcripts by IVM oocytes is particularly interesting, therefore, a variety of approaches were employed to determine whether the observed transcriptional changes during IVM were real or an artifact of the techniques used during analysis, including the analysis of transcript abundance in oocytes in vitro matured in the presence of a-amanitin. Subsets of the differentially expressed genes were also validated by quantitative real-time PCR (qPCR) and the gene expression data was classified according to gene ontology and pathway enrichment. Numerous cell cycle linked (CDC2, CDK5, CDK8, HSPA2, MAPK14, TXNL4B), molecular transport (STX5, STX17, SEC22A, SEC22B), and differentiation (NACA) related genes were found to be among the several over-expressed transcripts in GV oocytes compared to the matured counterparts, while ANXA1, PLAU, STC1and LUM were among the over-expressed genes after oocyte maturation. Conclusion: Using sequential experiments, we have shown and confirmed transcriptional changes during oocyte maturation. This dataset provides a unique reference resource for studies concerned with the molecular mechanisms controlling oocyte meiotic maturation in cattle, addresses the existing conflicting issue of transcription during meiotic maturation and contributes to the global goal of improving assisted reproductive technology.

Chemical potential and the nature of dark energy: The case of a phantom field

The influence of a possible nonzero chemical potential mu on the nature of dark energy is investigated by assuming that the dark energy is a relativistic perfect simple fluid obeying the equation of state, p=omega rho (omega < 0, constant). The entropy condition, S >= 0, implies that the possible values of omega are heavily dependent on the magnitude, as well as on the sign of the chemical potential. For mu > 0, the omega parameter must be greater than -1 (vacuum is forbidden) while for mu < 0 not only the vacuum but even a phantomlike behavior (omega <-1) is allowed. In any case, the ratio between the chemical potential and temperature remains constant, that is, mu/T=mu(0)/T(0). Assuming that the dark energy constituents have either a bosonic or fermionic nature, the general form of the spectrum is also proposed. For bosons mu is always negative and the extended Wien's law allows only a dark component with omega <-1/2, which includes vacuum and the phantomlike cases. The same happens in the fermionic branch for mu < 0. However, fermionic particles with mu > 0 are permitted only if -1

Large change in the predicted number of small halos due to a small amplitude oscillating inflaton potential

A smooth inflaton potential is generally assumed when calculating the primordial power spectrum, implicitly assuming that a very small oscillation in the inflaton potential creates a negligible change in the predicted halo mass function. We show that this is not true. We find that a small oscillating perturbation in the inflaton potential in the slow-roll regime can alter significantly the predicted number of small halos. A class of models derived from supergravity theories gives rise to inflaton potentials with a large number of steps and many trans-Planckian effects may generate oscillations in the primordial power spectrum. The potentials we study are the simple quadratic (chaotic inflation) potential with superimposed small oscillations for small field values. Without leaving the slow-roll regime, we find that for a wide choice of parameters, the predicted number of halos change appreciably. For the oscillations beginning in the 10(7)-10(8) M(circle dot) range, for example, we find that only a 5% change in the amplitude of the chaotic potential causes a 50% suppression of the number of halos for masses between 10(7)-10(8) M(circle dot) and an increase in the number of halos for masses <10(6) M(circle dot) by factors similar to 15-50. We suggest that this might be a solution to the problem of the lack of observed dwarf galaxies in the range 10(7)-10(8) M(circle dot). This might also be a solution to the reionization problem where a very large number of Population III stars in low mass halos are required.

Age distribution of the central stars of galactic disk planetary nebulae

Context. Determination of the ages of central stars of planetary nebulae (CSPN) is a complex problem, and there is presently no single method that can be generally applied. We have developed several methods of estimating the ages of CSPN, based on both the observed nebular properties and some properties of the stars themselves. Aims. Our aim is to estimate the ages and the age distribution of CSPN and to compare the derived results with mass and age determinations of CSPN and white dwarfs based on empirical determinations of these quantities. Methods. We considered a sample of planetary nebulae in the galactic disk, most of which (similar to 69%) are located in the solar neighbourhood, within 3 kpc from the Sun. We discuss several methods of deriving the age distribution of CSPN, namely; (i) the use of an age-metallicity relation that also depends on the galactocentric distance; (ii) the use of an age-metallicity relation obtained for the galactic disk; and (iii) the determination of ages from the central star masses obtained from the observed nitrogen abundances. Results. We estimated the age distribution of CSPN with average uncertainties of 1-2 Gyr, and compared our results with the expected distribution based both on the observed mass distribution of white dwarfs and on the age distribution derived from available mass distributions of CSPN. Based on our derived age distributions, we conclude that most CSPN in the galactic disk have ages under 6 Gyr, and that the age distribution is peaked around 2-4 Gyr.

MASS DISTRIBUTION IN HICKSON COMPACT GROUPS OF GALAXIES

This study presents the mass distribution for a sample of 18 late-type galaxies in nine Hickson compact groups. We used Ha rotation curves (RCs) from high-resolution two-dimensional velocity fields of Fabry-Perot observations and the J-band photometry from the Two Micron All Sky Survey, in order to determine the dark halo and the visible matter distributions. The study compares two halo density profiles, an isothermal core-like distribution, and a cuspy one. We also compare their visible and dark matter distributions with those of galaxies belonging to cluster and field galaxies coming from two samples: 40 cluster galaxies of Barnes et al. and 35 field galaxies of Spano et al. The central halo surface density is found to be constant with respect to the total absolute magnitude similar to what is found for the isolated galaxies. This suggests that the halo density is independent of galaxy type and environment. We have found that core-like density profiles better fit the RCs than cuspy-like ones. No major differences have been found between field, cluster, and compact group galaxies with respect to their dark halo density profiles.

An analytical approach to the dwarf galaxies cusp problem

Aims. An analytical solution for the discrepancy between observed core-like profiles and predicted cusp profiles in dark matter halos is studied. Methods. We calculate the distribution function for Navarro-Frenk-White halos and extract energy from the distribution, taking into account the effects of baryonic physics processes. Results. We show with a simple argument that we can reproduce the evolution of a cusp to a flat density profile by a decrease of the initial potential energy.

Galaxy distribution and evolution around a sample of 2dF groups

Context. We study galaxy evolution and spatial patterns in the surroundings of a sample of 2dF groups. Aims. Our aim is to find evidence of galaxy evolution and clustering out to 10 times the virial radius of the groups and so redefine their properties according to the spatial patterns in the fields and relate them to galaxy evolution. Methods. Group members and interlopers were redefined after the identification of gaps in the redshift distribution. We then used exploratory spatial statistics based on the the second moment of the Ripley function to probe the anisotropy in the galaxy distribution around the groups. Results. We found an important anticorrelation between anisotropy around groups and the fraction of early-type galaxies in these fields. Our results illustrate how the dynamical state of galaxy groups can be ascertained by the systematic study of their neighborhoods. This is an important achievement, since the correct estimate of the extent to which galaxies are affected by the group environment and follow large-scale filamentary structure is relevant to understanding the process of galaxy clustering and evolution in the Universe.

Beyond the Fragmentation Threshold Hypothesis: Regime Shifts in Biodiversity Across Fragmented Landscapes

Ecological systems are vulnerable to irreversible change when key system properties are pushed over thresholds, resulting in the loss of resilience and the precipitation of a regime shift. Perhaps the most important of such properties in human-modified landscapes is the total amount of remnant native vegetation. In a seminal study Andren proposed the existence of a fragmentation threshold in the total amount of remnant vegetation, below which landscape-scale connectivity is eroded and local species richness and abundance become dependent on patch size. Despite the fact that species patch-area effects have been a mainstay of conservation science there has yet to be a robust empirical evaluation of this hypothesis. Here we present and test a new conceptual model describing the mechanisms and consequences of biodiversity change in fragmented landscapes, identifying the fragmentation threshold as a first step in a positive feedback mechanism that has the capacity to impair ecological resilience, and drive a regime shift in biodiversity. The model considers that local extinction risk is defined by patch size, and immigration rates by landscape vegetation cover, and that the recovery from local species losses depends upon the landscape species pool. Using a unique dataset on the distribution of non-volant small mammals across replicate landscapes in the Atlantic forest of Brazil, we found strong evidence for our model predictions - that patch-area effects are evident only at intermediate levels of total forest cover, where landscape diversity is still high and opportunities for enhancing biodiversity through local management are greatest. Furthermore, high levels of forest loss can push native biota through an extinction filter, and result in the abrupt, landscape-wide loss of forest-specialist taxa, ecological resilience and management effectiveness. The proposed model links hitherto distinct theoretical approaches within a single framework, providing a powerful tool for analysing the potential effectiveness of management interventions.

Hormone-regulated expression and distribution of versican in mouse uterine tissues

Background: Remodeling of the extracellular matrix is one of the most striking features observed in the uterus during the estrous cycle and after hormone replacement. Versican (VER) is a hyaluronan-binding proteoglycan that undergoes RNA alternative splicing, generating four distinct isoforms. This study analyzed the synthesis and distribution of VER in mouse uterine tissues during the estrous cycle, in ovariectomized (OVX) animals and after 17beta-estradiol (E2) and medroxyprogesterone (MPA) treatments, either alone or in combination. Methods: Uteri from mice in all phases of the estrous cycle, and animals subjected to ovariectomy and hormone replacement were collected for immunoperoxidase staining for versican, as well as PCR and quantitative Real Time PCR. Results: In diestrus and proestrus, VER was exclusively expressed in the endometrial stroma. In estrus and metaestrus, VER was present in both endometrial stroma and myometrium. In OVX mice, VER immunoreaction was abolished in all uterine tissues. VER expression was restored by E2, MPA and E2+MPA treatments. Real Time PCR analysis showed that VER expression increases considerably in the MPA-treated group. Analysis of mRNA identified isoforms V0, V1 and V3 in the mouse uterus. Conclusion: These results show that the expression of versican in uterine tissues is modulated by ovarian steroid hormones, in a tissue-specific manner. VER is induced in the myometrium exclusively by E2, whereas MPA induces VER deposition only in the endometrial stroma.

Mechanisms of Vascular Damage by Hemorrhagic Snake Venom Metalloproteinases: Tissue Distribution and In Situ Hydrolysis

Background: Envenoming by viper snakes constitutes an important public health problem in Brazil and other developing countries. Local hemorrhage is an important symptom of these accidents and is correlated with the action of snake venom metalloproteinases (SVMPs). The degradation of vascular basement membrane has been proposed as a key event for the capillary vessel disruption. However, SVMPs that present similar catalytic activity towards extracellular matrix proteins differ in their hemorrhagic activity, suggesting that other mechanisms might be contributing to the accumulation of SVMPs at the snakebite area allowing capillary disruption. Methodology/Principal Findings: In this work, we compared the tissue distribution and degradation of extracellular matrix proteins induced by jararhagin (highly hemorrhagic SVMP) and BnP1 (weakly hemorrhagic SVMP) using the mouse skin as experimental model. Jararhagin induced strong hemorrhage accompanied by hydrolysis of collagen fibers in the hypodermis and a marked degradation of type IV collagen at the vascular basement membrane. In contrast, BnP1 induced only a mild hemorrhage and did not disrupt collagen fibers or type IV collagen. Injection of Alexa488-labeled jararhagin revealed fluorescent staining around capillary vessels and co-localization with basement membrane type IV collagen. The same distribution pattern was detected with jararhagin-C (disintegrin-like/cysteine-rich domains of jararhagin). In opposition, BnP1 did not accumulate in the tissues. Conclusions/Significance: These results show a particular tissue distribution of hemorrhagic toxins accumulating at the basement membrane. This probably occurs through binding to collagens, which are drastically hydrolyzed at the sites of hemorrhagic lesions. Toxin accumulation near blood vessels explains enhanced catalysis of basement membrane components, resulting in the strong hemorrhagic activity of SVMPs. This is a novel mechanism that underlies the difference between hemorrhagic and non-hemorrhagic SVMPs, improving the understanding of snakebite pathology.

Characterization of subgraph relationships and distribution in complex networks

A network can be analyzed at different topological scales, ranging from single nodes to motifs, communities, up to the complete structure. We propose a novel approach which extends from single nodes to the whole network level by considering non-overlapping subgraphs (i.e. connected components) and their interrelationships and distribution through the network. Though such subgraphs can be completely general, our methodology focuses on the cases in which the nodes of these subgraphs share some special feature, such as being critical for the proper operation of the network. The methodology of subgraph characterization involves two main aspects: (i) the generation of histograms of subgraph sizes and distances between subgraphs and (ii) a merging algorithm, developed to assess the relevance of nodes outside subgraphs by progressively merging subgraphs until the whole network is covered. The latter procedure complements the histograms by taking into account the nodes lying between subgraphs, as well as the relevance of these nodes to the overall subgraph interconnectivity. Experiments were carried out using four types of network models and five instances of real-world networks, in order to illustrate how subgraph characterization can help complementing complex network-based studies.