The trophic role of the squid Loligo plei as a keystone species in the South Brazil Bight ecosystem

The issue of whether loliginid squid can influence the average structure of marine ecosystems in a keystone role, i.e. a strong effect with relatively low biomass, has not yet been examined. Here, the diet of Loligo plei in inner shelf waters of the South Brazil Bight was examined, as a first step, based on the stomach contents of 2200 squid hand-jigged in shallow water (, 30 m) and taken as bycatch of shrimp trawlers in deeper water (30-100 m). Diet varied by size, season, and fishing zone. Stomachs were not empty in similar to 12%, with more empty during winter. The range of mantle lengths of squid caught by jigging (101-356 mm) appeared to differ from the squid trawled (30-236 mm), and the diet also differed. Food categories recorded in deeper water did not include amphipods or polychaetes, but in both fishing areas, fish were the most common prey. The fish prey identified included Trachurus lathami, small pelagic species, trichiurids, and Merluccius hubbsi. Demersal species, such as Ctenosciaena gracilicirrhus, and flatfish were also present. An ecosystem network model is updated through which a mixed-trophic impact matrix and ""keystoneness"" indicators were calculated. Loligo plei represents an important link between pelagic and demersal energy pathways, with high indices of keystoneness.

Plastic pellets as oviposition site and means of dispersal for the ocean-skater insect Halobates

Microplastics are omnipresent in the oceans and generally have negative impacts on the biota. However, flotsam may increase the availability of hard substrates, which are considered a limiting resource for some oceanic species, e.g. as oviposition sites for the ocean insect Halobates. This study describes the use of plastic pellets as an oviposition site for Halobates micans and discusses possible effects on its abundance and dispersion. Inspection of egg masses on stranded particles on beaches revealed that a mean of 24% (from 0% to 62%) of the pellets bore eggs (mean of 5 and max. of 48 eggs per pellet). Most eggs (63%) contained embryos, while 37% were empty egg shells. This shows that even small plastic particles are used as oviposition site by H. micans, and that marine litter may have a positive effect over the abundance and dispersion of this species. (C) 2012 Elsevier Ltd. All rights reserved.

Influence of dental materials used for sealing caries lesions on laser fluorescence measurements

The aim of this study was to determine the influence of thickness and aging on the intrinsic fluorescence of sealing materials and their ability to block fluorescence from the underlying surface as assessed using a laser fluorescence device. Cavities of 0.5 mm and 1 mm depth were drilled into acrylic boards which were placed over two surfaces with different fluorescence properties: a low-fluorescence surface, to assess the intrinsic fluorescence of the sealing materials, and a high-fluorescence surface, to assess the fluorescence-blocking ability of the sealing materials. Ten cavities of each depth were filled with different sealing materials: Adper Scotchbond Multi-Purpose, Adper Single Bond 2, FluroShield, Conseal f and UltraSeal XT Plus. Fluorescence was measured with a DIAGNOdent pen at five different time points: empty cavity, after polymerization, and 1 day, 1 week and 1 month after filling. The individual values after polymerization, as well as the area under the curve for the different periods were submitted to ANOVA and the Tukey test (p < 0.05). At 0.5 mm, Scotchbond, FluroShield and UltraSeal showed insignificant changes in intrinsic fluorescence with aging and lower fluorescence after polymerization than Single Bond and Conseal. At 1 mm, Scotchbond and FluroShield showed the lowest intrinsic fluorescence, but only Scotchbond showed no chagnes in fluorescence with aging. At both depths, Scotchbond blocked significantly less fluorescence. All sealing materials blocked more fluorescence when applied to a depth of 1 mm. At 0.5 mm, fissure sealants blocked more fluorescence than adhesives, and did not show significant changes with aging. Scotchbond had the least affect on the fluorescence from the underlying surface and would probably have the least affect on the monitoring of sealed dental caries by laser fluorescence.

Histomorphometric analysis of inflammatory response and necrosis in re-implanted central incisor of rats treated with low-level laser therapy

Low-level laser therapy is a tool employed in the management of post-operative inflammation process and in the enhancement of reparative process. The aim of the study was to perform histological evaluation of dental and periodontal ligament of rats central upper-left incisor teeth re-implanted and irradiated with low-level laser (InGaAl, 685 nm, 50 J/cm(2)) 15, 30, and 60 days after re-implantation. Seventy-two male rats had the central upper left incisor removed and kept for 15 min on dry gauze before replantation. Laser was irradiated over the root surface and empty alveolus prior replantation and over surrounding mucosa after the re-implantation. After histological procedures, all slices were analyzed regarding external resorption area and histological aspects. We observed an increase of root resorption (p < 0.05) in the control group compared to the laser group at 15, 30, and 60 days. These results showed that the laser groups developed less root resorption areas than the control group in all experimental periods. Additionally, histological analysis revealed less inflammatory cells and necrotic areas in laser groups.

Study of quercetin-loaded liposomes as potential drug carriers: in vitro evaluation of human complement activation

Liposomes have been employed as potential drug carriers. However, after their in vivo administration, they can be destabilized by proteins of complement system, contributing to the clearance of vesicles from blood circulation. Antioxidant flavonoids such as quercetin have been reported to be beneficial to human health, but their low water solubility and bioavailability limit their enteric administration. Therefore, the development of appropriate flavonoid-carriers could be of great importance to drug therapy. The aim of the present study was to evaluate the activation of human complement system proteins by liposomes composed of soya phosphatidylcholine (SPC) and cholesterol (CHOL) or cholesteryl ethyl ether (CHOL-OET) loaded with quercetin or not. The consumption of complement, via classical (CP) and alternative (AP) pathways, by different vesicles was evaluated using a hemolytic assay and quantitative determination of iC3b and natural antibodies deposited on empty liposomal surfaces by ELISA. The main results showed that empty liposomes composed of large amounts of CHOL consumed more complement components than the others for both CP and AP. Furthermore, replacement of CHOL with CHOL-OET reduced complement consumption via both CP and AP. Incorporation of quercetin did not change CP and AP consumption. Deposition of iC3b, IgG and IgM in vesicles composed of SPC: CHOL-OET at a molar ratio of 1.5:1 was lower compared to the others. Taken together, these observations suggest that liposomes composed of SPC: CHOL-OET at a molar ratio of 1.5:1 are the most appropriate among the vesicles studied herein to be used as a drug carrier system in further investigations.

Inactivating Mutations of the Human Luteinizing Hormone Receptor in Both Sexes

The human luteinizing hormone/chorionic gonadotropin receptor (LHCGR) plays a fundamental role in male and female reproductive physiology. Over the past 15 years, several homozygous or compound heterozygous loss-of-function mutations in the LHCGR gene have been described in males and females. In genetic males, mutations in LHCGR were associated with distinct degrees of impairment in pre- and postnatal testosterone secretion resulting in a phenotypic spectrum. Patients with the severe form of LH resistance have predominantly female external genitalia and absence of secondary sex differentiation at puberty. Patients with milder forms have predominantly male external genitalia with micropenis and/or hypospadias or only infertility without ambiguity. The undermasculization is associated with low basal, as well as human CG-stimulated, testosterone levels and elevated LH levels after pubertal age, without abnormal step-up in testosterone biosynthesis precursors. The testes have only slightly reduced size but mature Leydig cells are absent or scarce (Leydig cell hypoplasia). Genetic females with inactivating LHCGR mutations have female external genitalia, spontaneous breast and pubic hair development at puberty, and normal or late menarche followed by oligoamenorrhea and infertility. Estradiol and progesterone levels are normal for the early to midfollicular phase, but do not reach ovulatory or luteal phase levels. Serum LH levels are high whereas follicle-stimulating hormone levels are normal or only slightly increased. Pelvic ultrasound has demonstrated a small or normal uterus and normal or enlarged ovaries with cysts. The inactivating mutations of the LHCGR have provided important insights into distinct physiological roles of LH in reproduction of both sexes.

CHARACTERIZATION OF LYOPHILIZED LIPOSOMES PRODUCED WITH NON-PURIFIED SOY LECITHIN: A CASE STUDY OF CASEIN HYDROLYSATE MICROENCAPSULATION

A commercial casein hydrolysate was microencapsulated in liposomes produced with non-purified soy lecithin, cryoprotected with two different disaccharides and lyophilized. The encapsulation efficiency of casein hydrolysate ranged from 30 to 40%. The powders were analyzed by differential scanning calorimetry (DSC), scanning electron micrography (SEM), infrared spectroscopy (FTIR) and wide angle X-ray diffraction (WAXD). DSC data revealed the presence of an exothermal transition in empty lyophilized liposomes, which was ascribed to the presence of a quasicrystalline lamellar phase (intermediary characteristics between the L-beta and L-c phases). The addition of peptides to the liposomal system caused the disappearance of this exothermic phenomenon, as they were located in the polar headgroup portion of the bilayer, causing disorder and preventing the formation of the quasicrystalline phase. Infrared data indicated the presence of the peptides in the lyophilized formulations and showed that the cryoprotectants interacted effectively with the polar heads of phospholipids in the bilayer.

How far is C-0(Gamma, X) with Gamma discrete from C-0(K, X) spaces?

For a locally compact Hausdorff space K and a Banach space X we denote by C-0(K, X) the space of X-valued continuous functions on K which vanish at infinity, provided with the supremum norm. Let n be a positive integer, Gamma an infinite set with the discrete topology, and X a Banach space having non-trivial cotype. We first prove that if the nth derived set of K is not empty, then the Banach-Mazur distance between C-0(Gamma, X) and C-0(K, X) is greater than or equal to 2n + 1. We also show that the Banach-Mazur distance between C-0(N, X) and C([1, omega(n)k], X) is exactly 2n + 1, for any positive integers n and k. These results extend and provide a vector-valued version of some 1970 Cambern theorems, concerning the cases where n = 1 and X is the scalar field.

A method for harvesting unfermented pollen from stingless bees (Hymenoptera, Apidae, Meliponini)

Pollen traps used for harvesting pollen from Apis mellifera do not work for stingless bees, as most species have small entrances and rapidly deposit large quantities of propolis at any barrier in front of the nest. Some stingless beekeepers harvest pollen by removing it directly from pollen pots, but this pollen is normally fermented and unpalatable. The aim of this study was to test a new method for harvesting pollen from stingless bee colonies before it begins to ferment. Colonies of Scaptotrigona depilis were removed and replaced by empty hives, which were occupied by the returning foragers and used for storing pollen and nectar. After one week, the pollen and honey were harvested directly from the storing pots and weighed. On average, the colonies produced 8.7 g of honey and 54.2 g of unfermented pollen (n = 10). This method is a viable option for harvesting unfermented pollen from stingless bees, especially with species that harvest large amounts of pollen. The unfermented pollen of S. depilis was well received in taste tests, receiving higher scores than fermented pollen, and similar scores to A. mellifera pollen, so could have great commercial possibilities. It is also a good method for studying the foraging of stingless bees because the amount of harvested food can be easily and precisely quantified.

Large Mammals in an Agroforestry Mosaic in the Brazilian Atlantic Forest

The forest-like characteristics of agroforestry systems create a unique opportunity to combine agricultural production with biodiversity conservation in human-modified tropical landscapes. The cacao-growing region in southern Bahia, Brazil, encompasses Atlantic forest remnants and large extensions of agroforests, locally known as cabrucas, and harbors several endemic large mammals. Based on the differences between cabrucas and forests, we hypothesized that: (1) non-native and non-arboreal mammals are more frequent, whereas exclusively arboreal and hunted mammals are less frequent in cabrucas than forests; (2) the two systems differ in mammal assemblage structure, but not in species richness; and (3) mammal assemblage structure is more variable among cabrucas than forests. We used camera-traps to sample mammals in nine pairs of cabruca-forest sites. The high conservation value of agroforests was supported by the presence of species of conservation concern in cabrucas, and similar species richness and composition between forests and cabrucas. Arboreal species were less frequently recorded, however, and a non-native and a terrestrial species adapted to open environments (Cerdocyon thous) were more frequently recorded in cabrucas. Factors that may overestimate the conservation value of cabrucas are: the high proportion of total forest cover in the study landscape, the impoverishment of large mammal fauna in forest, and uncertainty about the long-term maintenance of agroforestry systems. Our results highlight the importance of agroforests and forest remnants for providing connectivity in human-modified tropical forest landscapes, and the importance of controlling hunting and dogs to increase the value of agroforestry mosaics.

Head-to-Head Comparison of Three Vaccination Strategies Based on DNA and Raw Insect-Derived Recombinant Proteins against Leishmania

Parasitic diseases plague billions of people among the poorest, killing millions annually, and causing additional millions of disability-adjusted life years lost. Leishmaniases affect more than 12 million people, with over 350 million people at risk. There is an urgent need for efficacious and cheap vaccines and treatments against visceral leishmaniasis (VL), its most severe form. Several vaccination strategies have been proposed but to date no head-to-head comparison was undertaken to assess which is the best in a clinical model of the disease. We simultaneously assayed three vaccination strategies against VL in the hamster model, using KMPII, TRYP, LACK, and PAPLE22 vaccine candidate antigens. Four groups of hamsters were immunized using the following approaches: 1) raw extracts of baculovirus-infected Trichoplusia ni larvae expressing individually one of the four recombinant proteins (PROT); 2) naked pVAX1 plasmids carrying the four genes individually (DNA); 3) a heterologous prime-boost (HPB) strategy involving DNA followed by PROT (DNA-PROT); and 4) a Control including empty pVAX1 plasmid followed by raw extract of wild-type baculovirus-infected T. ni larvae. Hamsters were challenged with L. infantum promastigotes and maintained for 20 weeks. While PROT vaccine was not protective, DNA vaccination achieved protection in spleen. Only DNA-PROT vaccination induced significant NO production by macrophages, accompanied by a significant parasitological protection in spleen and blood. Thus, the DNA-PROT strategy elicits strong immune responses and high parasitological protection in the clinical model of VL, better than its corresponding naked DNA or protein versions. Furthermore, we show that naked DNA coupled with raw recombinant proteins produced in insect larvae biofactories -the cheapest way of producing DNA-PROT vaccines-is a practical and cost-effective way for potential "off the shelf" supplying vaccines at very low prices for the protection against leishmaniases, and possibly against other parasitic diseases affecting the poorest of the poor.

DNA vaccine containing the mycobacterial hsp65 gene prevented insulitis in MLD-STZ diabetes

Abstract Background Our group previously demonstrated that a DNA plasmid encoding the mycobacterial 65-kDa heat shock protein (DNA-HSP65) displayed prophylactic and therapeutic effect in a mice model for tuberculosis. This protection was attributed to induction of a strong cellular immunity against HSP65. As specific immunity to HSP60 family has been detected in arthritis, multiple sclerosis and diabetes, the vaccination procedure with DNA-HSP65 could induce a cross-reactive immune response that could trigger or worsen these autoimmune diseases. Methods In this investigation was evaluated the effect of a previous vaccination with DNA-HSP65 on diabetes development induced by Streptozotocin (STZ). C57BL/6 mice received three vaccine doses or the corresponding empty vector and were then injected with multiple low doses of STZ. Results DNA-HSP65 vaccination protected mice from STZ induced insulitis and this was associated with higher production of IL-10 in spleen and also in the islets. This protective effect was also concomitant with the appearance of a regulatory cell population in the spleen and a decreased infiltration of the islets by T CD8+ lymphocytes. The vector (DNAv) also determined immunomodulation but its protective effect against insulitis was very discrete. Conclusion The data presented in this study encourages a further investigation in the regulatory potential of the DNA-HSP65 construct. Our findings have important implications for the development of new immune therapy strategies to combat autoimmune diseases.

Tissue distribution of DNA-Hsp65/TDM-loaded PLGA microspheres and uptake by phagocytic cells

This study aimed to demonstrate that microspheres, used as delivery vehicle of DNA-Hsp65/TDM [plasmid DNA encoding heat shock protein 65 (Hsp65) coencapsulated with trehalose dimycolate (TDM) into PLGA microspheres], are widely spread among several organs after intramuscular administration in BALB/c mice. In general, we showed that these particles were phagocytosed by antigen presenting cells, such as macrophages and dendritic cells. Besides, it was demonstrated herein that draining lymph node cells presented a significant increase in the number of cells expressing costimulatory molecules (CD80 and CD86) and MHC class II, and also that the administration of the DNA-Hsp65/TDM and vector/TDM formulations resulted in the up-regulation of CD80, CD86 and MHC class II expression when compared to control formulations (vector/TDM and empty). Regarding the intracellular trafficking we observed that following phagocytosis, the microspheres were not found in the late endosomes and/or lysosomes, until 15 days after internalization, and we suggest that these constructions were hydrolysed in early compartments. Overall, these data expand our knowledge on PLGA [poly (lactic-co- glycolic acid)] microspheres as gene carriers in vaccination strategies, as well as open perspectives for their potential use in clinical practice.

Eruption sequestrum - case report and histopathological findings

Eruption sequestrum is an uncommon disturbance in eruption and consists of small fragments of calcified tissue overlying the crowns of erupting permanent molar teeth, especially at the time of eruption of the mandibular first molars. This paper reports a case of unilateral eruption sequestrum in a 7-year-old Brazilian boy and describes its histopathological findings. A white small fragment, 0.5 cm in diameter, with hard consistency, irregular shape and located on the occlusal surface of the erupting mandibular left first molar was excised. Microscopic examination revealed large trabeculae with empty lacunae and a minimal amount of existing spongy bone consisting of acute inflammatory cells (neutrophils). Signs of necrosis were found on the periphery. The histological analysis was consistent with non-vital bone and the diagnosis of eruption sequestrum was established. Clinical and radiographic follow-up visits scheduled at short intervals and then every 6 months revealed normal postoperative conditions.

Characterization of lyophilized liposomes produced with non-purified soy lecithin: a case study of casein hydrolysate microencapsulation

A commercial casein hydrolysate was microencapsulated in liposomes produced with non-purified soy lecithin, cryoprotected with two different disaccharides and lyophilized. The encapsulation efficiency of casein hydrolysate ranged from 30 to 40%. The powders were analyzed by differential scanning calorimetry (DSC), scanning electron micrography (SEM), infrared spectroscopy (FTIR) and wide angle X-ray diffraction (WAXD). DSC data revealed the presence of an exothermal transition in empty lyophilized liposomes, which was ascribed to the presence of a quasicrystalline lamellar phase (intermediary characteristics between the Lβ and Lc phases). The addition of peptides to the liposomal system caused the disappearance of this exothermic phenomenon, as they were located in the polar headgroup portion of the bilayer, causing disorder and preventing the formation of the quasicrystalline phase. Infrared data indicated the presence of the peptides in the lyophilized formulations and showed that the cryoprotectants interacted effectively with the polar heads of phospholipids in the bilayer.