Increase of leishmanicidal and tubercular activities using steroids linked to aminoquinoline

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cultivo de Haematococcus pluvialis Flotow (Chlorophyceae) em sistema fototrófico e mixotrófico a base de meio alternativo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efficacy of Oral and Parenteral Routes of Mycobacterium bovis Bacille Calmette-Guerin Vaccination Against Experimental Bovine Tuberculosis in White-Tailed Deer (Odocoileus virginianus): A Feasibility Study

We investigated the efficacy of oral and parenteral Mycobacterium bovis bacille Calmette-Guerin Danish strain 1331 (BCG) in its ability to protect white-tailed deer (Odocoileus virginianus) against disease caused by M. bovis infection. Twenty-two white-tailed deer were divided into four groups. One group (n=5) received 109 colony-forming units (cfu) BCG via a lipid-formulated oral bait; one group (n=5) received 109 cfu BCG in culture directly to the oropharynx, one group (n=6) was vaccinated with 106 cfu BCG subcutaneously, and one group served as a control and received culture media directly to the oropharynx (n=6). All animals were challenged 3 mo after vaccination. Five months postchallenge the animals were examined for lesions. Results indicate that both oral forms of BCG and parenterally administerd BCG offered significant protection against M. bovis challenge as compared to controls. This study suggests that oral BCG vaccination may be a feasible means of controlling bovine tuberculosis in wild white-tailed deer populations.

Detection Methods for the Genus Lysobacter and the Species Lysobacter enzymogenes

Strains of Lysobacter enzymogenes, a bacterial species with biocontrol activity, have been detected via 16S rDNA sequences in soil in different parts of the world. In most instances, however, their occurrence could not be confirmed by isolation, presumably because the species occurred in low numbers relative to faster-growing species of Bacillus or Pseudomonas. In this study, we developed DNA-based detection and enrichment culturing methods for Lysobacter spp. and L. enzymogenes specifically. In the DNA-based method, a region of 16S rDNA conserved among Lysobacter spp. (L4: GAG CCG ACG TCG GAT TAG CTA GTT), was used as the forward primer in PCR amplification. When L4 and universal bacterial primer 1525R were used to amplify DNA from various bacterial species, an 1100-bp product was found in Lysobacter spp. exclusively. The enrichment culturing method involved culturing soils for 3 days in a chitin-containing broth amended with antibiotics. Bacterial strains in the enrichment culture were isolated on yeast-cell agar and then identified by 16S rDNA sequence analysis. A strain of L. enzymogenes added to soils was detected at populations as low as 102 and 104 CFU/g soil by PCR amplification and enrichment culturing, respectively. In a survey of 58 soil samples, Lysobacter was detected in 41 samples by PCR and enrichment culture, out of which 6 yielded strains of Lysobacter spp. by enrichment culture. Among isolated strains, all were identified to be L. enzymogenes, with the exception of a strain of L. antibioticus. Although neither method alone is completely effective at detecting L. enzymogenes, they are complementary when used together and may provide new information on the spatial distribution of the species in soil.

Parental Ethnotheories of Child Development: Looking Beyond Independence and Individualism in American Belief Systems

Over the past several decades, the topic of child development in a cultural context has received a great deal of theoretical and empirical investigation. Investigators from the fields of indigenous and cultural psychology have argued that childhood is socially and historically constructed, rather than a universal process with a standard sequence of developmental stages or descriptions. As a result, many psychologists have become doubtful that any stage theory of cognitive or socialemotional development can be found to be valid for all times and places. In placing more theoretical emphasis on contextual processes, they define culture as a complex system of common symbolic action patterns (or scripts) built up through everyday human social interaction by means of which individuals create common meanings and in terms of which they organize experience. Researchers understand culture to be organized and coherent, but not homogenous or static, and realize that the complex dynamic system of culture constantly undergoes transformation as participants (adults and children) negotiate and re-negotiate meanings through social interaction. These negotiations and transactions give rise to unceasing heterogeneity and variability in how different individuals and groups of individuals interpret values and meanings. However, while many psychologists—both inside and outside the fields of indigenous and cultural psychology–are now willing to give up the idea of a universal path of child development and a universal story of parenting, they have not necessarily foreclosed on the possibility of discovering and describing some universal processes that underlie socialization and development-in-context. The roots of such universalities would lie in the biological aspects of child development, in the evolutionary processes of adaptation, and in the unique symbolic and problem-solving capacities of the human organism as a culture-bearing species. For instance, according to functionalist psychological anthropologists, shared (cultural) processes surround the developing child and promote in the long view the survival of families and groups if they are to demonstrate continuity in the face of ecological change and resource competition, (e.g. Edwards & Whiting, 2004; Gallimore, Goldenberg, & Weisner, 1993; LeVine, Dixon, LeVine, Richman, Leiderman, Keefer, & Brazelton, 1994; LeVine, Miller, & West, 1988; Weisner, 1996, 2002; Whiting & Edwards, 1988; Whiting & Whiting, 1980). As LeVine and colleagues (1994) state: A population tends to share an environment, symbol systems for encoding it, and organizations and codes of conduct for adapting to it (emphasis added). It is through the enactment of these population-specific codes of conduct in locally organized practices that human adaptation occurs. Human adaptation, in other words, is largely attributable to the operation of specific social organizations (e.g. families, communities, empires) following culturally prescribed scripts (normative models) in subsistence, reproduction, and other domains [communication and social regulation]. (p. 12) It follows, then, that in seeking to understand child development in a cultural context, psychologists need to support collaborative and interdisciplinary developmental science that crosses international borders. Such research can advance cross-cultural psychology, cultural psychology, and indigenous psychology, understood as three sub-disciplines composed of scientists who frequently communicate and debate with one another and mutually inform one another’s research programs. For example, to turn to parental belief systems, the particular topic of this chapter, it is clear that collaborative international studies are needed to support the goal of crosscultural psychologists for findings that go beyond simply describing cultural differences in parental beliefs. Comparative researchers need to shed light on whether parental beliefs are (or are not) systematically related to differences in child outcomes; and they need meta-analyses and reviews to explore between- and within-culture variations in parental beliefs, with a focus on issues of social change (Saraswathi, 2000). Likewise, collaborative research programs can foster the goals of indigenous psychology and cultural psychology and lay out valid descriptions of individual development in their particular cultural contexts and the processes, principles, and critical concepts needed for defining, analyzing, and predicting outcomes of child development-in-context. The project described in this chapter is based on an approach that integrates elements of comparative methodology to serve the aim of describing particular scenarios of child development in unique contexts. The research team of cultural insiders and outsiders allows for a look at American belief systems based on a dialogue of multiple perspectives.

Macrofaunal succession in sediments around kelp and wood falls in the deep NE Pacific and community overlap with other reducing habitats

Sunken parcels of macroalgae and wood provide important oases of organic enrichment at the deep-sea floor, yet sediment community structure and succession around these habitat islands are poorly evaluated. We experimentally implanted 100-kg kelp falls and 200 kg wood falls at 1670 m depth in the Santa Cruz Basin to investigate (1) macrofaunal succession and (2) species overlap with nearby whale-fall and cold-seep communities over time scales of 0.25-5.5 yr. The abundance of infaunal macrobenthos was highly elevated after 0.25 and 0.5 yr near kelp parcels with decreased macrofaunal diversity and evenness within 0.5 m of the falls. Apparently opportunistic species (e.g., two new species of cumaceans) and sulfide tolerant microbial grazers (dorvilleid polychaetes) abounded after 0.25-0.5 yr. At wood falls, opportunistic cumaceans become abundant after 0.5 yr, but sulfide tolerant species only became abundant after 1.8-5.5 yr, in accordance with the much slower buildup of porewater sulfides at wood parcels compared with kelp falls. Species diversity decreased significantly over time in sediments adjacent to the wood parcels, most likely due to stress resulting from intense organic loading of nearby sediments (up to 20-30% organic carbon). Dorvilleid and ampharetid polychaetes were among the top-ranked fauna at wood parcels after 3.0-5.5 yr. Sediments around kelp and wood parcels provided low-intensity reducing conditions that sustain a limited chemoautrotrophically-based fauna. As a result, macrobenthic species overlap among kelp, wood, and other chemosynthetic habitats in the deep NE Pacific are primarily restricted to apparently sulfide tolerant species such as dorvilleid polychaetes, opportunistic cumaceans, and juvenile stages of chemosymbiont containing vesicomyid bivalves. We conclude that organically enriched sediments around wood falls may provide important habitat islands for the persistence and evolution of species dependent on organic- and sulfide-rich conditions at the deep-sea floor and contribute to beta and gamma diversity in deep-sea ecosystems. (C) 2010 Elsevier Ltd. All rights reserved.

Accidents by external causes in adolescents: care in sentinel urgency and emergency services in the Brazilian State Capitals-2009

Adolescents are seeking new references and experiences, which may involve attitudes of risk and exposure to accidents and violence from external causes. These events constitute a serious Public Health problem. The scope of this study was to analyze the occurrence of accidents by external causes in adolescents from 10 to 19 years of age attended at sentinel urgency and emergency services in Brazil. Data from the 2009 Surveillance System for Violence and Accidents (VIVA 2009) was analyzed in 74 emergency units in 23 state capitals and the Federal District. The findings revealed that 6,434 adolescents (89.8%) were victims of accidents and 730 (10.2 %) were victims of violence. The main causes of the accidents were falls and traffic accidents, and assaults were predominant in violence. For both accidents and violence, non-white male adolescents were predominant and the events occurred most frequently on the public highways. A marked increase was detected, with hospitalization of victims of violence between 15 and 19 years of age. Understanding the epidemiological reality of external causes among adolescents represents an important tool for health prevention and promotion policies and the culture of peace seeking to reduce morbidity and mortality.

Maize and soybeans production in integrated system under no-tillage with different pasture combinations and animal categories

The adoption of no-till system (NTS) combined with crop-livestock integration (CLI) has been a strategy promoted in Brazil, aiming to maximize areas yield and increase agribusiness profitability. This study aimed to evaluate grains yield and phytotechnical attributes from maize and soybean culture by CLI system under NTS after winter annual pure and diversified pastures with the absence or presence of grazing animals. The experiment was installed in Castro (Parana State, Brazil) on in a dystrophic Humic Rhodic Hapludox with a clay texture, using experimental design of randomized complete blocks in 4 x 2 factorial scheme with three replications. Treatments included four pasture combinations (diversified or pure) and animal categories (light and heavy) subjected or not to grazing animals during the winter. During 2008/09 and 2009/10 summers, the area was cultivated with soybeans and maize, respectively, with yield assessment of grains and phytotechnical attributes. Treatments did not alter the yield and weight of a thousand seeds (WTS) of soybeans. In maize culture, the grazing animal during the winter increased the plant population and grains yield, but gave slight decrease in WTS. Pasture combinations (diversified or pure) and animal categories (light and heavy) did not interfere in soybean culture, but benefited the maize crop.

Regeneration and characterization of somatic hybrids combining sweet orange and mandarin/mandarin hybrid cultivars for citrus scion improvement

Protoplast fusion between sweet orange and mandarin/mandarin hybrids scion cultivars was performed following the model "diploid embryogenic callus protoplast + diploid mesophyll-derived protoplast". Protoplasts were isolated from embryogenic calli of 'Pera' and 'Westin' sweet orange cultivars (Citrus sinensis) and from young leaves of 'Fremont', Nules', and 'Thomas' mandarins (C. reticulata), and 'Nova' tangelo [C. reticulata x (C. paradisi x C. reticulata)]. The regenerated plants were characterized based on their leaf morphology (thickness), ploidy level, and simple sequence repeat (SSR) molecular markers. Plants were successfully generated only when 'Pera' sweet orange was used as the embryogenic parent. Fifteen plants were regenerated being 7 tetraploid and 8 diploid. Based on SSR molecular markers analyses all 7 tetraploid regenerated plants revealed to be allotetraploids (somatic hybrids), including 2 from the combination of 'Pera' sweet orange + 'Fremont' mandarin, 3 'Pera' sweet orange + 'Nules' mandarin, and 2 'Pera' sweet orange + 'Nova' tangelo, and all the diploid regenerated plants showed the 'Pera' sweet orange marker profile. Somatic hybrids were inoculated with Alternaria alternata and no disease symptoms were detected 96 h post-inoculation. This hybrid material has the potential to be used as a tetraploid parent in interploid crosses for citrus scion breeding.

Effect of supplementation of two sources and two levels of copper on lipid metabolism in Nellore beef cattle

This study was conducted with 35 Nellore beef cattle to determine the effect of supplementation of two levels and two copper sources (organic and inorganic) on metabolism of lipids and cholesterol of meat. The five treatments used were: Control: without copper supplementation, 110 or 140: 10 or 40 mg/kg DM (as Cu sulfate), O10 or O40: 10 or 40 mg/kg DM (as Cu proteinate). In general, the copper supplementation changed the fatty acid profile of meat (p < 0.05), with a higher proportion of unsaturated fatty acids and reduction of saturated fatty acids. There was no effect of supplementation on blood cholesterol and triglycerides, however; in general, there was a reduction in cholesterol concentration in the L dorsi (p < 0.05) compared to the control treatment through the reduction (p < 0.05) in the concentrations of GSH and GSH/GSSG ratio. The Cu supplementation did have an influence on metabolism of lipids. The production of healthier meat is beneficial to public health by reducing the risk of cardiovascular disease. (C) 2012 Elsevier Ltd. All rights reserved.

Anatomical and ultrastructural analyses of in vitro organogenesis from root explants of commercial passion fruit (Passiflora edulis Sims)

This study aimed to characterize the anatomical events and ultrastructural aspects of direct and indirect in vitro organogenesis in Passiflora edulis. Root explants were cultured on induction medium, supplemented with 4.44 mu M 6-benzyladenine. Roots at different stages of development were collected and processed for observation by light microscopy and scanning and transmission electron microscopy. Patterns of direct and indirect regeneration were observed in the explants. During direct organogenesis, the organogenic buds and nodules, formed from meristemoids, originated from the pericycle regions distant from the cut surface. Completely differentiated buds were observed after 20 days of culture. During indirect organogenesis, bud formation occurred via meristemoids at the periphery of the calli, which differentiated from the cortical region of the initial explant. Regardless of the regeneration pattern, the meristemoids had similar ultrastructural characteristics; however, differences were reported in the nuclear shape of the cells of the meristemoids formed directly and indirectly. This study provides important information for enhancing the understanding and characterization of the organogenic process in non-meristematic explants and provides information on the use of roots as explants in genetic transformation protocols for this important tropical species.

In Vitro Development and Cell Allocation After Aggregation of Syngeneic Wild Type and Fluorescence-Expressing Bovine Cloned Embryos

Background: The in vitro production (IVP) of embryos by in vitro fertilization or cloning procedures has been known to cause epigenetic changes in the conceptus that in turn are associated with abnormalities in pre- and postnatal development. Handmade cloning (HMC) procedures and the culture of zona-free embryos in individual microwells provide excellent tools for studies in developmental biology, since embryo development and cell allocation patterns can be evaluated under a wide range of embryo reconstruction arrangements and in in vitro embryo culture conditions. As disturbances in embryonic cell allocation after in vitro embryo manipulations and unusual in vivo conditions during the first third of pregnancy appear to be associated with large offspring, embryo aggregation procedures may allow a compensation for epigenetic defects between aggregated embryos or even may influence more favorable cell allocation in embryonic lineages, favoring subsequent development. Thus, the aim of this study was to evaluate in vitro embryo developmental potential and the pattern of cell allocation in blastocysts developed after the aggregation of handmade cloned embryos produced using syngeneic wild type and/or transgenic somatic cells. Materials, Methods & Results: In vitro-matured bovine cumulus-oocyte complexes (COC) were manually bisected after cumulus and zona pellucida removal; then, two enucleated hemi-oocytes were paired and fused with either a wild type (WT) or a GFP-expressing (GFP) fetal skin cell at the 11th and 19th passages, respectively. Following chemical activation, reconstructed cloned embryos and zona-free parthenote embryos were in vitro-cultured in microwells, for 7 days, either individually (1 x 100%) or after the aggregation of two structures (2 x 100%) per microwell, as follows: (G1) one WT cloned embryo; (G2) two aggregated WT embryos; (G3) one GFP cloned embryo; (G4) two aggregated GFP embryos; (G5) aggregation of a WT embryo and a GFP embryo; (G6) one parthenote embryo; or (G7) two aggregated parthenote embryos. Fusion (clones), cleavage (Day 2), and blastocyst (Day 7) rates, and embryonic cell allocation were compared by the. 2 or Fisher tests. Total cell number (TCN) in blastocysts was analyzed by the Student's test (P < 0.05). Fusion and cleavage rates, and cell allocation were similar between groups. On a per WOW basis, development to the blastocyst stage was similar between groups, except for lower rates of development seen in G3. However, when based on number of embryos per group (one or two), blastocyst development was higher in G1 than all other groups, which were similar between one another. Cloned GFP embryos had lower in vitro development to the blastocyst stage than WT embryos, which had more TCN than parthenote or aggregated chimeric WT/GFP embryos. Aggregated GFP embryos had fewer cells than the other embryo groups. Discussion: The in vitro development of GFP cloned embryos was lower than WT embryos, with no effects on cell allocation in resulting blastocysts. Differences in blastocyst rate between groups were likely due to lower GFP-expressing cell viability, as GFP donor cells were at high population cell doublings when used for cloning. On a per embryo basis, embryo aggregation on Day 1 resulted in blastocyst development similar to non-aggregated embryos on Day 7, with no differences in cell proportion between groups. The use of GFP-expressing cells was proven a promising strategy for the study of cell allocation during embryo development, which may assist in the elucidation of mechanisms of abnormalities after in vitro embryo manipulations, leading to the development of improved protocols for the in vitro production (IVP) of bovine embryos.

Malakoplakia after renal transplantation in the current era of immunosuppressive therapy: case report and literature review

Malakoplakia is a rare chronic granulomatous disease of unknown cause. It is thought to be caused by an acquired bactericidal defect of macrophages. Malakoplakia is associated with chronic infections and immunosuppression. Although it occurs mainly in the urinary tract, it has already been reported in almost every organ system. The isolation of bacteria, especially Escherichia coli, is common in malakoplakia patients. Here, we present a case of primary cutaneous malakoplakia in a kidney transplant recipient who had been taking prednisone, tacrolimus, and mycophenolate. Culture of a lesion grew Burkholderia cepacia complex. Treatment with high doses of trimethoprim-sulfamethoxazole was successful. We also present a systematic review of the literature, identifying 4 previously reported cases of malakoplakia after renal transplantation under similar immunosuppressive therapy, most occurring in the urinary tract or perineum and following benign courses to cure. Data in the literature suggest that malakoplakia has become even rarer since changes were made in the immunosuppressive therapy employed after kidney transplantation.

HEPATOTOXICITY ASSESSMENT OF THE AZO DYES DISPERSE ORANGE 1 (DO1), DISPERSE RED 1 (DR1,) AND DISPERSE RED 13 (DR13) IN HEPG2 CELLS

During the dyeing process in baths approximately 10 to 15% of the dyes used are lost and reach industrial effluents, thus polluting the environment. Studies showed that some classes of dyes, mainly azo dyes and their by-products, exert adverse effects on humans and local biota, since the wastewater treatment systems and water treatment plants were found to be ineffective in removing the color and reducing toxicity of some dyes. In the present study, the toxicity of the azo dyes disperse orange 1 (DO1), disperse red 1 (DR1), and disperse red 13 (DR13) was evaluated in HepG2 cells grown in monolayers or in three dimensional (3D) culture. Hepatotoxicity of the dyes was measured using 3-(4,5-dimethylthiazol-2yl)2,5-diphenyltetrazolium (MTT) and cell counting kit 8 (CCK-8) assays after 24, 48, and 72 h of incubation of cells with 3 different concentrations of the azo dyes. The dye DO1 only reduced the mitochondrial activity in HepG2 cells grown in a monolayer after 72 h incubation, while the dye DR1 showed this deleterious effect in both monolayer and 3D culture. In contrast, dye DR13 decreased the mitochondrial activity after 24, 48, and 72 h of exposure in both monolayer and 3D culture. With respect to dehydrogenase activity, only the dye DR13 diminished the activity of this enzyme after 72 h of exposure in both monolayer and 3D culture. Our results clearly demonstrated that exposure to the studied dyes induced cytotoxicity in HepG2 cells.

Biochemical and metabolic profiles of Trichoderma strains isolated from common bean crops in the Brazilian Cerrado, and potential antagonism against Sclerotinia sclerotiorum

Some species of Trichoderma have successfully been used in the commercial biological control of fungal pathogens, e.g., Sclerotinia sclerotiorum, an economically important pathogen of common beans (Phaseolus vulgaris L.). The objectives of the present study were (1) to provide molecular characterization of Trichoderma strains isolated from the Brazilian Cerrado; (2) to assess the metabolic profile of each strain by means of Biolog FF Microplates; and (3) to evaluate the ability of each strain to antagonize S. sclerotiorum via the production of cell wall-degrading enzymes (CWDEs), volatile antibiotics, and dual-culture tests. Among 21 isolates, we identified 42.86 % as Trichoderma asperellum, 33.33 % as Trichoderma harzianum, 14.29 % as Trichoderma tomentosum, 4.76 % as Trichoderma koningiopsis, and 4.76 % as Trichoderma erinaceum. Trichoderma asperellum showed the highest CWDE activity. However, no species secreted a specific group of CWDEs. Trichoderma asperellum 364/01, T. asperellum 483/02, and T. asperellum 356/02 exhibited high and medium specific activities for key enzymes in the mycoparasitic process, but a low capacity for antagonism. We observed no significant correlation between CWDE and antagonism, or between metabolic profile and antagonism. The diversity of Trichoderma species, and in particular of T. harzianum, was clearly reflected in their metabolic profiles. Our findings indicate that the selection of Trichoderma candidates for biological control should be based primarily on the environmental fitness of competitive isolates and the target pathogen. (C) 2012 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.