The double of the doubles of Klein surfaces

A Klein surface is a surface with a dianalytic structure. A double of a Klein surface $X$ is a Klein surface $Y$ such that there is a degree two morphism (of Klein surfaces) $Y\rightarrow X$. There are many doubles of a given Klein surface and among them the so-called natural doubles which are: the complex double, the Schottky double and the orienting double. We prove that if $X$ is a non-orientable Klein surface with non-empty boundary, the three natural doubles, although distinct Klein surfaces, share a common double: "the double of doubles" denoted by $DX$. We describe how to use the double of doubles in the study of both moduli spaces and automorphisms of Klein surfaces. Furthermore, we show that the morphism from $DX$ to $X$ is not given by the action of an isometry group on classical surfaces.Comment: 14 pages; more details in the proof of theorem

Abnormal regulation of Na,K-ATPase in Glucose Intolerant Rats.

Introduction: Glucose is the most important physiological insulin secretagogue. However, the mechanisms underlying glucose-induced insulin release are not fully understood. The role of electrogenic systems such as ionic pumps, to these events remains essentially uninvestigated. Na,K-ATPase, responsible for maintaining Na+ and K+ gradients across the plasma membrane and generates a net outward current, thus changes in its activity may contribute to the early ionic events regulating insulin secretion (Therien and Blostein, 2000). Objective: The aim of this work was to evaluate the regulation of Na,K-ATPase activity by glucose in intact -cells of normal and glucose intolerant (GI) rats and its putative contribution to the regulation of insulin secretion. Material and Methods: Pancreatic -cells, from normal or control or GI rats, were isolated and cultured (48h). Cell batches were pre-incubated (30min) with 2mM glucose to reach basal. Afterwards cells were challenged with glucose in the interval 0-11mM for 60min, for dose-dependence evaluation, or with 8mM glucose for 5-120min, for time-dependence evaluation. ATPase activity was assessed in intact cells by colorimetric quantification of Pi formed in 30min. Na,K-ATPase activity was calculated by the difference between the activities obtained in the absence and in presence the of 1mM ouabain (Costa et al., 2009). Results: In β-cells from normal rats, glucose induced a bimodal regulation of Na,K-ATPase. In the absence of glucose, Na,K-ATPase activity was 0.056±0.015 U/mg. Stimulation with 2mM glucose induced an increase of Na,K-ATPase activity of ~4 fold whereas for [glucose] above 2mM it was observed a significant inhibition of Na,K-ATPase activity (0.061±0.013, 0.080±0.009 and 0.064±0.005 U/mg for 5.6, 8.4 and 11mM glucose, respectively, compared to 0.188±0.035 U/mg observed in 2mM G; n=3-8). β-cells from GI rats does not present this profile; in the absence of glucose, Na,K-ATPase activity was 0.202±0.036 U/mg and no significant differences from this value were observed with the other glucose concentration tested. Addicionally, in β-cells from normal rats, glucose (8mM) induced a time-dependent inhibition, with a biphasic profile, of Na,K-ATPase - it was observed a decrease in the pump activity between 0 and 20min stimulation where it reached a minimum value (77%). For incubation periods over 20min, the pump activity slowly and partially recovered (54%, 55% and 52%, for 30, 60 and 120min, respectively; n=7). In β-cells from GI animals, an less accentuated decrease of Na,K-ATPase activity between 0 ans 20min was also observed (34%), and is not observed further recover in activity. Conclusions: This work demonstrates there Na,K-ATPase is strictly regulated by glucose in pancreatic β-cell. This regulation is unpaired in GI animals. Na,K-ATPase contribution to glucose-induced ionic events and insulin secretion might be relevant and must be explored as a possible therapeutic target in TD2 . 1. Therien AG, Blostein R (2000) Mechanisms of sodium pump regulation. Am J Physiol Cell Physiol 279:C541-C566 2. Costa AR, Real J, Antunes CM, Cruz-Morais J (2009) A new approach for determination of Na,K-ATPase activity: application to intact pancreatic beta-cells. In Vitro Cell Dev Biol Ani

Implication of AMPK in glucose-evoked modulation of Na,K-ATPase

Background and aims: Na,K-ATPase is an integral membrane protein that maintains the gradients of Na+ and K+, using the energy of ATP hydrolysis, maintaining the ionic gradients that allow electrical activity to occur. It has been demonstrated that, in pancreatic β-cells, Na,K-ATPase is regulated by glucose and that this phenomenon is impaired in glucose intolerant subjects. However, the mechanism underlying glucose-induced modulation of Na,K-ATPase is still unclear. The AMP-activated protein kinase (AMPK) is a molecular key player in energy homeostasis, providing exquisite sensitivity to small changes in intracellular AMP levels and thus to intracellular [ATP]/[ADP] ratio, that is known to activate protein regulatory pathways. Since in pancreatic β-cell, glucose has marked effects on oxidative metabolism and total intracellular ATP and AMP levels, the involvement of AMPK in the cascade of events regulating Na,K-ATPase regulation in pancreatic β-cells was postulated. The aim of this work was to evaluate the putative role of AMPK in the glucose-evoked regulation of Na,K-ATPase activity in the pancreatic β-cell. Materials and methods: Pancreatic -cells from normal (control) or glucose-intolerant Wistar rats (GIR) were isolated and cultured (48h). Cell batches were pre-incubated (30min) with 2.1mM glucose to reach basal activity. Afterwards cells were challenged to 8.4mM glucose for 20min, in the presence or absence of AMPK agonists (AICAR) and antagonists (compound C; CC). ATPase activity was assessed in intact cells by colorimetric quantification of Pi formed in 30min. Na,K-ATPase activity was calculated by the difference between the activities obtained in the absence and in presence the of 1mM ouabain. Results: In basal conditions the activity of Na,K-ATPase from normal and GIR pancreatic β-cell was similar (0.184±0.030 and 0.186±0.020 molPi/min/mgProt, respectively). Challenging the control β-cells with glucose 8.4mM evoked a 62% reduction of Na,K-ATPase activity whereas in GIR β-cells a significantly lower inhibition (40%) was observed. The addition of AICAR 1mM abolished glucose-induced Na,K-ATPase inhibition (0,166±0.011 molPi/min/mg). In control β-cell, the addition of CC 10 μM had no effect on glucose-induced inhibition of Na,K-ATPase. In the contrary, in GIR β-cells it significantly potentiated glucose-evoked inhibition of Na,K-ATPase reaching values similar to that observed in the controls (66%). Conclusions: The AMPK agonist AICAR counteracts the inhibitory action of glucose on Na,K-ATPase of control β-cells whereas CC amplified the glucose-induced inhibition of Na,K-ATPase in GIR β-cells. These results suggest that AMPK plays a central role in the cascade of events underlying glucose-induced modulation of Na,K-ATPase and that the defect must be upstream of AMPK. Finally, abnormal glucose-induced regulation of Na,K-ATPase occurs prior to overt type 2 diabetes and might be a feature in the disease development

An integrative assessment to determine the genotoxic hazard of estuarine sediments: combining cell and whole-organism responses

The application of the Comet assay in environmental monitoring remains challenging in face of the complexity of environmental stressors, e.g., when dealing with estuarine sediments, that hampers the drawing of cause-effect relationships. Although the in vitro Comet assay may circumvent confounding factors, its application in environmental risk assessment (ERA) still needs validation. As such, the present work aims at integrating genotoxicity and oxidative DNA damage induced by sediment-bound toxicants in HepG2 cells with oxidative stress-related effects observed in three species collected from an impacted estuary. Distinct patterns were observed in cells exposed to crude mixtures of sediment contaminants from the urban/industrial area comparatively to the ones from the rural/riverine area of the estuary, with respect to oxidative DNA damage and oxidative DNA damage. The extracts obtained with the most polar solvent and the crude extracts caused the most significant oxidative DNA damage in HepG2 cells, as measured by the formamidopyrimidine-DNA glycosylase (FPG)-modified Comet assay. This observation suggests that metals and unknown toxicants more hydrophilic than polycyclic aromatic hydrocarbons may be important causative agents, especially in samples from the rural part of the estuary, where oxidative DNA damage was the most significant. Clams, sole, and cuttlefish responded differentially to environmental agents triggering oxidative stress, albeit yielding results accordant with the oxidative DNA damage observed in HepG2 cells. Overall, the integration of in vivo biomarker responses and Comet assay data in HepG2 cells yielded a comparable pattern, indicating that the in vitro FPG-modified Comet assay may be an effective and complementary line-of-evidence in ERA even in particularly challenging, natural, scenarios such as estuarine environments

Judicial Risk and Credit Market Performance: Micro Evidence from Brazil Payroll Loans

A large body of literature has stressed the institution-development nexus as critical in explaining differences in countries' economic performance. The empirical evidence, however, has been mainly at the aggregate level, associating macro performance with measures of quality of institutions. This paper, by relating a judicial decision on the legality of payroll debit loans in Brazil to bank-level decision variables, provides micro evidence on how creditor legal protection affects market performance. Payroll loans are personal loans with principal and interests payments directly deducted from the borrowers' payroll check, which, in practice, makes a collateral out of future income. In June 2004, a high-level federal court upheld a regional court ruling that had declared payroll deduction illegal. Using personal loans without payroll deduction as a control group, we assess whether the ruling had an impact on market performance. Evidence indicates that it had an adverse impact on risk perception, interest rates, and amount lent.

Environmental risk assessment in a contaminated estuary: an integrated weight of evidence approach as a decision support tool

Environmental risk assessment of complex ecosystems such as estuaries is a challenge, where innovative and integrated approaches are needed. The present work aimed at developing an innovative integrative methodology to evaluate in an impacted estuary (the Sado, in Portugal, was taken as case study), the adverse effects onto both ecosystem and human health. For the purpose, new standardized lines of evidence based on multiple quantitative data were integrated into a weight of evidence according to a best expert judgment approach. The best professional judgment for a weight of evidence approach in the present study was based on the following lines of evidence: i) human contamination pathways; ii) human health effects: chronic disease; iii) human health effects: reproductive health; iv) human health effects: health care; v) human exposure through consumption of local agriculture produce; vi) exposure to contaminated of water wells and agriculture soils; vii) contamination of the estuarine sedimentary environment (metal and organic contaminants); viii) effects on benthic organisms with commercial value; and ix) genotoxic potential of sediments. Each line of evidence was then ordinally ranked by levels of ecological or human health risk, according to a tabular decision matrix and expert judgment. Fifteen experts scored two fishing areas of the Sado estuary and a control estuarine area, in a scale of increasing environmental risk and management actions to be taken. The integrated assessment allowed concluding that the estuary should not be regarded as impacted by a specific toxicant, such as metals and organic compounds hitherto measured, but by the cumulative risk of a complex mixture of contaminants. The proven adverse effects on species with commercial value may be used to witness the environmental quality of the estuarine ecosystem. This method argues in favor of expert judgment and qualitative assessment as a decision support tool to the integrative management of estuaries. Namely it allows communicating environmental risk and proposing mitigation measures to local authorities and population under a holistic perspective as an alternative to narrow single line of evidence approaches, which is mandatory to understand cause and effect relationships in complex areas like estuaries.info:eu-repo/semantics/publishedVersio

Biomarkers: a strategic tool in the assessment of environmental quality of coastal waters

Ecosystems are under the pressure of complex mixtures of contaminants whose effects are not always simple to assess. Biomarkers, acting as early warning signals of the presence of potentially toxic xenobiotics, are useful tools for assessing either exposure to, or the effects of these compounds providing information about the toxicant bioavailability. In fact, it has been argued that a full understanding of ecotoxicological processes must consider an integrated multi-level approach, in which molecular impact is related with higher-order biological consequences at the individual, population and community levels. Monitoring programs should make use of this tool to link contaminants and ecological responses fulfilling strategies like those launched by OSPAR (Commissions of Oslo and Paris) Convention on the protection of the marine environment of the North-East Atlantic and the International Council for the Exploration of the Sea (ICES). An overview of the work done in the past few years using biomarkers as in situ tools for pollution assessment in Portuguese coastal waters is presented as a contribution to the set up of a biomonitoring program for the Portuguese coastal zone. Considering the data set available the biomonitoring proposal should include the analysis of biomarkers and effects at individual levels. The aim of the program will include a spatial and temporal characterization of the biomarkers acetyl-cholinesterase, metallothioneins, DNA damage, adenylate energy charge and scope-for-growth levels. The investigation of the spatial variation of biomarkers is crucial to define sites for long term monitoring, which will be integrated with a chemical monitoring program. This framework will be a major contribution to the implementation of a national database for the use of biomarkers along the Portuguese coast.info:eu-repo/semantics/publishedVersio

Inhalable fucoidan microparticles combining two antitubercular drugs with potential application in pulmonary tuberculosis therapy

The pulmonary delivery of antitubercular drugs is a promising approach to treat lung tuberculosis. This strategy not only allows targeting the infected organ instantly, it can also reduce the systemic adverse effects of the antibiotics. In light of that, this work aimed at producing fucoidan-based inhalable microparticles that are able to associate a combination of two first-line antitubercular drugs in a single formulation. Fucoidan is a polysaccharide composed of chemical units that have been reported to be specifically recognised by alveolar macrophages (the hosts of Mycobacterium). Inhalable fucoidan microparticles were successfully produced, effectively associating isoniazid (97%) and rifabutin (95%) simultaneously. Furthermore, the produced microparticles presented adequate aerodynamic properties for pulmonary delivery with potential to reach the respiratory zone, with a mass median aerodynamic diameter (MMAD) between 3.6-3.9 mu m. The formulation evidenced no cytotoxic effects on lung epithelial cells (A549), although mild toxicity was observed on macrophage-differentiated THP-1 cells at the highest tested concentration (1 mg/mL). Fucoidan microparticles also exhibited a propensity to be captured by macrophages in a dose-dependent manner, as well as an ability to activate the target cells. Furthermore, drug-loaded microparticles effectively inhibited mycobacterial growth in vitro. Thus, the produced fucoidan microparticles are considered to hold potential as pulmonary delivery systems for the treatment of tuberculosis.Portuguese Foundation for Science and Technology [PTDC/DTP-FTO/0094/2012, UID/Multi/04326/2013, UID/BIM/04773/2013]; CAPES-Brazil [BEX 1168/13-4

Looking at the Trees in the Central Forest: A New Pallidal-Striatal Cell Type

The glopus pallidus is a central nucleus of the basal ganglia, pivotal to their function in health and disease. In this issue of Neuron, Mallet et al. (2012) reveal that this structure is more diverse than previously thought, and identify a novel cell type that projects from pallidum to striatum providing massive GABAergic innervation. These findings invite new views on basal ganglia processing