Proteoliposomes as matrix vesicles' biomimetics to study the initiation of skeletal mineralization

During the process of endochondral bone formation, chondrocytes and osteoblasts mineralize their extracellular matrix by promoting the formation of hydroxyapatite (HA) seed crystals in the sheltered interior of membrane-limited matrix vesicles (MVs). Ion transporters control the availability of phosphate and calcium needed for HA deposition. The lipidic microenvironment in which MV-associated enzymes and transporters function plays a crucial physiological role and must be taken into account when attempting to elucidate their interplay during the initiation of biomineralization. In this short mini-review, we discuss the potential use of proteoliposome systems as chondrocyte- and osteoblast-derived MVs biomimetics, as a means of reconstituting a phospholipid microenvironment in a manner that recapitulates the native functional MV microenvironment. Such a system can be used to elucidate the interplay of MV enzymes during catalysis of biomineralization substrates and in modulating in vitro calcification. As such, the enzymatic defects associated with disease-causing mutations in MV enzymes could be studied in an artificial vesicular environment that better mimics their in vivo biological milieu. These artificial systems could also be used for the screening of small molecule compounds able to modulate the activity of MV enzymes for potential therapeutic uses. Such a nanovesicular system could also prove useful for the repair/treatment of craniofacial and other skeletal defects and to facilitate the mineralization of titanium-based tooth implants.

The expression of melanopsin and clock genes in Xenopus laevis melanophores and their modulation by melatonin

Vertebrates have a central clock and also several peripheral clocks. Light responses might result from the integration of light signals by these clocks. The dermal melanophores of Xenopus laevis have a photoreceptor molecule denominated melanopsin (OPN4x). The mechanisms of the circadian clock involve positive and negative feedback. We hypothesize that these dermal melanophores also present peripheral clock characteristics. Using quantitative PCR, we analyzed the pattern of temporal expression of Opn4x and the clock genes Per1, Per2, Bmal1, and Clock in these cells, subjected to a 14-h light:10-h dark (14L:10D) regime or constant darkness (DD). Also, in view of the physiological role of melatonin in the dermal melanophores of X. laevis, we determined whether melatonin modulates the expression of these clock genes. These genes show a time-dependent expression pattern when these cells are exposed to 14L:10D, which differs from the pattern observed under DD. Cells kept in DD for 5 days exhibited overall increased mRNA expression for Opn4x and Clock, and a lower expression for Per1, Per2, and Bmal1. When the cells were kept in DD for 5 days and treated with melatonin for 1 h, 24 h before extraction, the mRNA levels tended to decrease for Opn4x and Clock, did not change for Bmal1, and increased for Per1 and Per2 at different Zeitgeber times (ZT). Although these data are limited to one-day data collection, and therefore preliminary, we suggest that the dermal melanophores of X. laevis might have some characteristics of a peripheral clock, and that melatonin modulates, to a certain extent, melanopsin and clock gene expression.

Citizens against the State: the riddle of high impact, low functionality courts in Brazil

The Brazilian federal judiciary offers an interesting riddle to scholars of judicial politics and policy change. While the courts have played a major policy role over the past two decades, constraining and altering federal policy across a range of subjects, the court system has simultaneously been labeled "dysfunctional." This paper investigates this riddle: a system plagued by major systemic flaws in its day-to-day operations, which nonetheless still manages to exert a powerful influence on public policy in Brazil. I adopt a new institutional perspective, focusing on how the institutional and normative structure within which judges and other legal actors operate affects policy outcomes.

Climate governance in an international system under conservative hegemony: the role of major powers

In the last five years, climate change has been established as a central civilizational driver of our time. As a result of this development, the most diversified social processes - as well as the fields of science which study them - have had their dynamics altered. In International Relations, this double challenge could be explained as follows: 1) in empirical terms, climate change imposes a deepening of cooperation levels on the international community, considering the global common character of the atmosphere; and 2) to International Relations as a discipline, climate change demands from the scientific community a conceptual review of the categories designed to approach the development of global climate governance. The goal of this article is to discuss in both conceptual and empirical terms the structure of global climate change governance, through an exploratory research, aiming at identifying the key elements that allow understanding its dynamics. To do so, we rely on the concept of climate powers. This discussion is grounded in the following framework: we now live in an international system under conservative hegemony that is unable to properly respond to the problems of interdependence, among which - and mainly -, the climate issue.

Nitric oxide: a major determinant of mast cell phenotype and function

Mast cells (MC) are important in the numerous physiological processes of homeostasis and disease. Most notably, MC are critical effectors in the development and exacerbation of allergic disorders. Nitric oxide (NO) is a diatomic radical produced by nitric oxide synthase (NOS), and has pluripotent cell signaling and cytotoxic properties. NO can influence many MC functions. Recent evidence shows the source of this NO can be from the mast cell itself. Governing the production of this endogenous NO, through alterations in the expression of tetrahydrobiopterin (BH4), a NOS cofactor, has stabilizing effects on MC degranulation. Furthermore, NO regulates the synthesis and secretion of de novo generated mediators, including leukotrienes and chemokines. These novel observations add to the growing body of knowledge surrounding the role of NO in the MC.

Sloth biology: an update on their physiological ecology, behavior and role as vectors of arthropods and arboviruses

This is a review of the research undertaken since 1971 on the behavior and physiological ecology of sloths. The animals exhibit numerous fascinating features. Sloth hair is extremely specialized for a wet tropical environment and contains symbiotic algae. Activity shows circadian and seasonal variation. Nutrients derived from the food, particularly in Bradypus, only barely match the requirements for energy expenditure. Sloths are hosts to a fascinating array of commensal and parasitic arthropods and are carriers of various arthropod-borne viruses. Sloths are known reservoirs of the flagellate protozoan which causes leishmaniasis in humans, and may also carry trypanosomes and the protozoan Pneumocystis carinii.

Between system maker and privileges taker: the role of China in the Greater Mekong Sub-region

Utilizing China's leadership projects in the Great Mekong Sub-Region (GMS) as a case study, this paper aims to investigate whether China qualifies as an international leader. This work argues that its geographic position and economic rise allow China to be a "system maker and privilege taker," which is a dual role forming in economic-political relations in the GMS in the last ten years. China is among major driving forces to set up an economic zone in GMS. Growing Chinese regional power is intimately related to the creation of various hubs connecting regional transportation, communication and energy systems that foster the economic development of this region. However, China also proves dark sides of rising powers which take advantage of their privileges to gain benefits. As a "system maker" with its own position and capability, China has notably benefited from building hydropower systems. More importantly, while China is pursuing its benefits and privileges, its hydropower projects have caused some negative effects for the ecosystem in the region. The inflation of dam constructions in both China and GMS countries is raising concerns about using natural resources of the Mekong River. Our concluding part addresses the pressing need to start a serious discussion on the balance between national interests and regional solidarity within the formulation of Chinese foreign policy in GMS.

Epidemiological role of humans, dogs and cats in the transmission of Trypanosoma cruzi in a central area of Argentina

Trypanosoma cruzi prevalence rates of human, dog and cat populations from 47 households of 3 rural localities of the phytogeographical Chaqueña area of Argentina were determined both by serological and xenodiagnostic procedures. Human prevalence rates were uniform and ranged from 49.6 to 58.7%. Overall prevalence rate in dogs (75.0%) was significantly higher than in humans (51.0%). The overall proportion of parasitemic individuals assessed by xenodiagnosis was significantly higher in either dog (64.2%) or cat (63.6%) populations than among humans (12.5%). Although both the average number of resident as well as infected individuals per household was higher for people than for dogs (6.5 vs. 3.3, and 3.4 vs. 2.4, respectively), the reverse was recorded when parasitemic individuals were considered (1.0 vs. 2.1). Results are discussed in relation to dog between dogs and people, and dogs and bugs. In the light of present data, dogs must be considered as the major donors of parasites to vector bugs and thus, principal contributors to transmission in this region of Argentina.

Bancroftian filariasis in two urban areas of Recife, Brasil: the role of individual risk factors

Bancroftian filariasis is spreading in towns of endemic areas as in Recife, northeastern Brazil, where it is a major public health problem. This paper deals with the prevalence of microfilaraemia and filarial disease in two urban areas of Recife, studying their association with individual characteristics and variables related to the exposure to the vectors. The parasitologic survey was performed through a "door-to-door" census and microfilaraemia was examined by the thick-drop technique using 45µl of peripheral blood collected between 20:00 and 24:00 o' clock. 2,863 individuals aged between 5 and 65 years were interviewed and submitted to clinical examination. Males aged between 15 and 44 years old presented the greatest risk of being microfilaraemic. Microfilaraemia was also significantly associated with no use of bednet to sleep. The risk of being microfilaraemic was greater among those who had lived in the studied areas for more than 5 years. The overall disease prevalence was 6.3%. Males presented the greatest risk of developing acute disease. The risk of developing chronic manifestations was also greater among males and increased with age. We found no association between time of residence, bednet use, microfilaraemia and acute and chronic disease. We may conclude that in endemic areas there are subgroups of individuals who has a higher risk of being microfilariae carriers due to different behaviours in relation to vector contact.

Mapping of a Leishmania major gene/locus that confers pentamidine resistance by deletion and insertion of transposable element

Pentamidine (PEN) is an alternative compound to treat antimony-resistant leishmaniasis patients, which cellular target remains unclear. One approach to the identification of prospective targets is to identify genes able to mediate PEN resistance following overexpression. Starting from a genomic library of transfected parasites bearing a multicopy episomal cosmid vector containing wild-type Leishmania major DNA, we isolated one locus capable to render PEN resistance to wild type cells after DNA transfection. In order to map this Leishmania locus, cosmid insert was deleted by two successive sets of partial digestion with restriction enzymes, followed by transfection into wild type cells, overexpression, induction and functional tests in the presence of PEN. To determine the Leishmania gene related to PEN resistance, nucleotide sequencing experiments were done through insertion of the transposon Mariner element of Drosophila melanogaster (mosK) into the deleted insert to work as primer island. Using general molecular techniques, we described here this method that permits a quickly identification of a functional gene facilitating nucleotide sequence experiments from large DNA fragments. Followed experiments revealed the presence of a P-Glycoprotein gene in this locus which role in Leishmania metabolism has now been analyzed.

Lysozyme plays a dual role against the dimorphic fungus Paracoccidioides brasiliensis

In order to determine the role of lysozyme, an antimicrobial peptide belonging to the innate immune system, against the dimorphic fungus Paracoccidioides brasiliensis, co-cultures of the MH-S murine alveolar macrophages cell line with P. brasiliensis conidia were done; assays to evaluate the effect of physiological and inflammatory concentrations of lysozyme directly on the fungus life cycle were also undertaken. We observed that TNF-α-activated macrophages significantly inhibited the conidia to yeast transition (p = 0.0043) and exerted an important fungicidal effect (p = 0.0044), killing 27% more fungal propagules in comparison with controls. Nonetheless, after adding a selective inhibitor of lysozyme, the fungicidal effect was reverted. When P. brasiliensis propagules were exposed directly to different concentrations of lysozyme, a dual effect was observed. Physiologic concentrations of the enzyme facilitated the conidia-to-yeast transition process (p < 0.05). On the contrary, inflammatory concentrations impaired the normal temperature-dependant fungal transition (p < 0.0001). When yeast cells were exposed to lysozyme, irrespective of concentration, the multiple-budding ability was badly impaired (p < 0.0001). In addition, ultra-structural changes such as subcellular degradation, fusion of lipid vacuoles, lamellar structures and interruption of the fibrilar layer were observed in lysozyme exposed conidia. These results suggest that lysozyme appears to exert a dual role as part of the anti-P. brasiliensis defense mechanisms.

Physiological responses of matrinxã (Brycon amazonicus) fed different levels of vitamin C and submitted to air exposure

The role of vitamin C on physiological responses of matrinxã (Brycon amazonicus) submitted to air exposure was analyzed. Nine hundred fish (70.15 g) were distributed in fifteen 500 l boxes (60 fish.box-1) and fed five rations (treatments): Control (no vitamin C); T100 (100 mg); T200 (200 mg); T400 (400 mg) and T800 (800 mg of vitamin C kg.ration-1). Each ration was offered to fish of three boxes during 60 days before the stress challenge that consisted of exposing fish to air for two minutes. Samplings were carried out for 5, 15, 30 and 60 minutes after the air exposure. Blood was collected for glucose, cortisol, total protein, sodium, chloride, hematocrit, hemoglobin determination, and white and red cell count. Liver was removed for hepatosomatic index (HSI) calculation and glycogen determination. Vitamin C did not affect the levels of cortisol, chloride, total protein, hemoglobin, leukocytes, hepatic glycogen or HSI in air exposed fish. Blood glucose levels elevation observed 60 minutes after the challenge did not depend on the levels of vitamin C, nor did the drop in serum sodium levels verified 60 minutes after stressor. In general, hematocrit did not change by effect of vitamin C but it was lower at 15 and 30 minutes after the challenge. The number of erythrocytes decreased in fish after 5 minute sampling in all treatments, especially at 30 and 60 minutes. The air exposure evoked alterations in stress indicators of matrinxã, and the vitamin C did not alter the responses.

Evolution of holometabola insect digestive systems: physiological and biochemical aspects

This review takes into account primarily the work done in our laboratory with insects from the major Holometabola orders. Only the most significant data for each insect will be presented and a proposal on the evolution of Holometabola insect digstive systems will be advanced.

Naturally acquired antibodies against the major merozoite surface coat protein (MSP-1) of Plasmodium falciparum acquired by residents in an endemic area of Colombia

A preliminary baseline epidemiological malaria survey was conducted in the village of Punta Soldado, Colombia. Parasite prevalence and density as well as serological data were obtained from 151 asymptomatic children and adults. Fifty individuals were infected with Plasmodium falciparum. The mean parasite density was 184 parasites/mm3. Greater than 90 of the sample population were P. falciparum antibody positive as detected by the indirect immunofluorescent antibody test (IFAT). The enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against the major merozoite surface protein (MSP-1) of P. falciparum. In this population, anti-MSP-1 antibody concentration is acquired in an age dependent manner with equal immunogenicity to both the N- and C-terminal regions of the molecule. Infection at the time of sampling was associated with a higher anti-MSP-1 antibody concentration than that found in non-infected individuals. Further studies are planned to assess the role of immune and non-immune factors in limiting the number of cases of severe malaria seen in this population.