Effects of past climate on Passiflora actinia (Passifloraceae) populations and insights into future species management in the Brazilian Atlantic forest

The Brazilian Atlantic Rainforest is one of the most diverse and threatened ecoregions on the planet and displays high levels of endemism. Despite several population analyses and phylogeographical studies, the origins of its species richness and the evolutionary processes that gave rise to this diversification remain poorly understood, especially at the southern edge of the Atlantic Forest. Passiflora actinia is an indigenous species from the southern Atlantic Forest and, as such, was influenced by climatic changes during the Pleistocene. In this study, we investigated the effects of past climate changes on the genetic diversity of P. actinia, using nuclear and plastid markers. We subsequently suggest strategies for the preservation of this species in particular and the whole ecoregion in general. We employed phylogeographical methods and combined these results with past, present and future ensemble niche models. Genetic variability in P. actinia was similar to that of other species with similar geographical distributions, and geographical structuring was not observed based on either type of genetic marker. Diversification in P. actinia was dated to the Pleistocene, suggesting that climate changes could have influenced the distribution of genetic diversity in this species. Our results predicted that suitable P. actinia habitat will persist in the highlands but will be reduced in the lowlands, especially with higher greenhouse gas concentrations. Conservation efforts should focus on populations with unique genetic units and populations from areas with greater climatic instability. Habitat loss due to deforestation in the Atlantic Forest constitutes a major risk to this species, especially to small populations or those with low diversity indices.     

Design of an enzyme cocktail consisting of different fungal platforms for efficient hydrolysis of sugarcane bagasse: Optimization and synergism studies

Lignocellulosic materials represent a very important and promising source of renewable biomass. In order to turn them into fermentable sugars, synergism among the different enzymes that carry out bioconversion of these materials is one of the main factors that should be considered. Experimental mixture design was performed to optimize the proportion of enzymes produced by native strains of Trichoderma harzianum IOC 3844, Penicillium funiculosum ATCC 11797, and Aspergillus niger ATCC 1004, resulting in a proportion of 15, 50, and 35%, respectively. This mixture was able to hydrolyze 25 g/L of pretreated sugarcane bagasse with 91% of yield after 48 h of enzymatic reaction. Synergism along the hydrolysis process, besides the influence of lignin, hemicellulose, and solids loading, were also studied. Response surface methodology (RSM) based on Central Composite Rotatable Design was used to optimize solids and protein loadings to increase glucose release and enzymatic hydrolysis yield. The optimum solid and protein loadings established with RSM were 196 g/L and 24 mg/g cellulose, respectively, and under these conditions (94.1 ± 8) g/L of glucose were obtained, corresponding to a hydrolysis yield of 64%. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1222–1229, 2016     

Genetic relationships among domesticated and wild sunflowers (Helianthus annuus,Asteraceae)

Randomly amplified polymorphic DNA (RAPD) loci were used to investigate the origin and genetic relationships of the domesticated sunflower and its wild relatives. A total of 13 primers was employed for the PCR amplifications, from which 68 polymorphic loci were scored. Analysis of RAPD data supports the origin of the domesticated sunflower from wildH. annuus. The high RAPD identity between wild and domesticatedH. annuus (I = 0.976 to I = 0.997) is concordant with a progenitorderivative relationship. However, the identities are very high and therefore provide little information regarding the geographic origin of the domesticated sunflower. Nonetheless, some inferences concerning relationships among domesticated sunflower accessions can be made. The native American varieties and old landracesform a genetically cohesive group based on RAPD evidence, probably due to their origin prior to the use of interspecific hybridization in the development of sunflower cultivars. In contrast, the modern cultivars are not genetically cohesive, perhaps due to the extensive use of intraspecific and interspecific hybridization in the development of modern sunflower varieties. Likewise, little concordance was observed between the geographical origin and genetic clustering of wild populations—an observation probably best explained by the weedy, human dispersed nature of wildH. annuus populations. The information presented here may be a reliable indicator of genetic relationships among wild and domesticated sunflower accessions. However, the processes generating the observed relationships are complex, and the occurrence of unexpected groupings or absence of predicted ones will probably remain difficult to understand.     

Population structuring of the ubiquitous stingless bee Tetragonisca angustula in southern Brazil as revealed by microsatellite and mitochondrial markers

Tetragonisca angustula is one of the most widespread stingless bees in the Neotropics. This species swarms frequently and is extremely successful in urban environments. In addition, it is one of the most popular stingless bee species for beekeeping in Latin America, so nest transportation and trading is common. Nest transportation can change the genetic structure of the host population, reducing inbreeding and increasing homogenization. Here, we evaluate the genetic structure of 17 geographic populations of T. angustula in southern Brazil to quantify the level of genetic differentiation between populations. Analyses were conducted on partially sequenced mitochondrial genes and 11 microsatellite loci of 1002 workers from 457 sites distributed on the mainland and on 3 islands. Our results show that T. angustula populations are highly differentiated as demonstrated by mitochondrial DNA (mtDNA) and microsatellite markers. Of 73 haplotypes, 67 were population‐specific. MtDNA diversity was low in 9 populations but microsatellite diversity was moderate to high in all populations. Microsatellite data suggest 10 genetic clusters and low level of gene flow throughout the studied area. However, physical barriers, such as rivers and mountain ranges, or the presence or absence of forest appear to be unrelated to population clusters. Factors such as low dispersal, different ecological conditions, and isolation by distance are most likely shaping the population structure of this species. Thus far, nest transportation has not influenced the general population structure in the studied area. However, due to the genetic structure we found, we recommend that nest transportation should only occur within and between populations that are genetically similar.     

Partial biochemical and immunochemical characterization of avian eggshell extracellular matrices.

There is evidence to suggest that extracellular matrix molecules, such as proteoglycans, are involved in the regulation of mineral deposition in calcifying tissues. One mineralizing system which is characterized by extremely rapid mineralization is the hen eggshell. This eggshell consists of a pair of nonmineralized eggshell membranes subjacent to the calcified eggshell proper; the eggshell proper is organized into palisades (columns) of mineralized matrix separated by pores. Between the membranes and the shell proper are compacted foci of tissue called mammillary knobs, which are thought to be sites where mineralization is initiated. Previous work from this laboratory has shown the presence of types I, V, and X collagen in the shell membranes. To address the question of the possible role of proteoglycans and glycosaminoglycans in mineralization of the eggshell, two approaches were used. First, immunohistochemistry was performed with monoclonal antibodies to various proteoglycan and glycosaminoglycan epitopes. This analysis indicates that different glycosaminoglycans are localized to discrete regions within the eggshell. Dermatan sulfate is present within the matrix of the shell proper and, to a lesser extent, the mammillary knobs and the outer portion of the shell membranes. In contrast, keratan sulfate is found in the shell membranes and prominently in the mammillary knobs. Interestingly, different keratan sulfate antibodies immunostain distinct regions of the eggshell, which suggests that various types of keratan sulfate are distributed differently. The second approach utilized was to extract the eggshell membranes and recover anionic molecules by anion-exchange chromatography. This resulted in the extraction of material which was recognized by antibodies to keratan sulfate, but not to chondroitin sulfate. This material was very large, as evidenced by its elution in the void volume of a Sepharose CL-2B column. The large size may be due to the extensive cross-links known to occur in the eggshell. If eggshell membranes are extracted at elevated temperature, the material recovered is of much smaller size. These results indicate that molecules recognized by antibodies to glycosaminoglycans are present in the eggshell, and their localized distribution relative to the calcified matrix suggests that they may be involved in the regulation of mineral deposition.     

Actinide complexes with the Kläui tripodal oxygen ligand - crystal structure of [{η-C5H5(CH2C(CH3)3)}Co{P(O)(OC2H5)2}3]2U

Reaction between [(C₅H₅)Co{P(O)(OEt)₂}₃]₂UCl₂ and neopentyl lithium affords the novel complex, [{η⁴-C₅H₅(CH₂C(CH₃)₃)}Co{P(O)(OEt)₂}₃]₂U, in which the uranium metal center has been dehalogenated and the neopentyl nucleophiles have attacked the cyclopentadienyl groups on the Kläui ([(C₅H₅)Co{P(O)(OEt)₂}₃]⁻) ligands. The uranium atom in the title compound possesses octahedral geometry defined by the oxygen atoms from two sets of tripodal oxygen ligands, while the cyclopentadienyl ligands are bound η⁴ to the cobalt atoms. The formation of this complex suggests that the Kläui ligand may not be a suitable ligand framework for supporting organometallic complexes of oxophilic early actinides.     

Evidence for the synthesis of thymosin alpha 1 by calf thymocytes and the production of this peptide by natural processing

Thymus and thymocytes from calf were extracted under isotonic conditions in the presence of protease inhibitors or under severe denaturing conditions (after quick freezing and thawing in boiling 0.1 M NaCl). The extracts, as well as the medium in which the thymocytes were obtained from thymus fragments (thymocyte supernatants), were size-fractionated by ultrafiltration. As in whole thymus isotonic extracts, thymosin alpha 1 [A. L. Goldstein, T. L. K. Low, M. McAdoo, J. McClure, G. B. Thurman, J. Rossio, C-Y. Lai, D. Chang, S-S. Wang, C. Harvey, A. H. Ramel, and J. Meienhofer (1977) Proc. Natl. Acad. Sci. USA 74, 725-729] was contained in isotonic extracts from thymocytes and also in thymocyte supernatants, as determined by isoelectric focusing and reverse-phase HPLC analysis. The extraction under denaturing conditions mainly yielded products with molecular masses over 50,000, showing very similar isoelectric focusing patterns in both thymocytes and whole thymus extracts. As deduced by isoelectric focusing analysis of diverse size-fractionated products, a strong association capacity seems to be responsible for an apparently high molecular mass of the components of these extracts. According to the pI, two of these components were prothymosin alpha [A. A. Haritos, G. J. Goodall, and B. L. Horecker (1984) Proc. Natl. Acad. Sci. USA 81, 1008-1011] and thymosin alpha 1. Prothymosin alpha was not detected in any isotonic extracts or thymocyte supernatants. These data suggest that calf thymocytes are capable of producing thymosin alpha 1, which would arise by natural processing of its precursor.     

Patterned cell adhesion associated with tissue deformations during dorsal closure in Drosophila

Cell shape changes within epithelia require the regulation of adhesive molecules that maintain tissue integrity. How remodelling of cell contacts is achieved while tissue integrity is maintained remains a fundamental question in morphogenesis. Dorsal Closure is a good system to study the dynamics of DE-Cadherin during morphogenesis. It relies on concerted cell shape changes of two epithelial sheets: amnioserosa cell contraction and epidermal cell elongation. To investigate the modulation of DE-Cadherin we performed antibody uptake experiments in live embryos during Dorsal Closure. We found that some antibodies access certain epitopes of the extracellular domain of native DE-Cadherin only in the amnioserosa and epidermal cells attached to the amnioserosa, which has never been observed in fixed DE-Cadherin in Drosophila embryos. These differences correlate with the different cell behaviour of these regions and therefore we suggest that DE-Cadherin exists in different forms that confer different adhesive strengths. We propose this to be a widespread mechanism for the differential modulation of adhesion during morphogenesis.     

Influence of diets supplemented with vitamins C and E on pirarucu (Arapaima gigas) blood parameters

This study evaluated the influence of diets supplemented with 500, 800, 1200 mg kg-1 of vitamin C (ascorbic acid or AA) and vitamin E (alpha-tocopherol or alpha-T) on the physiological responses of pirarucu fed for 2 months. Weight and mortality were not affected by dietary vitamin type or their concentrations. Significant increase (p<0.05) on the red blood cells count was obtained on treatments with 800 and 1200 mg AA kg-1 and on the hemoglobin concentration on treatment with 500 mg alpha-T kg-1 relatively to control. Mean corpuscular volume presented a significant decrease (p<0.05) on treatment with 800 and 1200 mg AA kg-1 when compared to control. Mean corpuscular hemoglobin concentration was significantly high (p<0.05) on treatment with 500 mg alpha-T kg-1. Only in vitamin C treatments, we noticed a significant increase (p<0.05) in the number of leucocytes relative to control. All fish in the vitamin-supplemented treatments, except 500 mg AA kg-1, had high total protein values compared to control. Fish treated with 800 or 1200 mg alpha-T kg-1 also showed increases in plasma glucose concentrations. Our results suggest that 800 and 1200 mg AA kg-1 are probably the most suitable concentrations for pirarucu diets, although high vitamin E diets are not necessary for quantitative leucocyte increases for this species.     

Análisis cladístico del género Pachycereus (Cactaceae) con caracteres morfológicos

El géneroPachycereus incluye 13 especies de acuerdo con las clasificaciones más recientes, y se distribuye en Guatemala, México y SW de los Estados Unidos, Se presenta un análisis filogenético con el propósito de fundamentar la monofilia del género, respecto a los otros géneros de la subtribu Pachycereinae. Se creó una matriz utilizando 44 caracteres estructurales (morfología y anatomía) y 29 taxones, incluyendo nueve especies de Pachycereinae. Los resultados indican que la subtribu Pachycereinae sólo puede ser monofilética si se incluyen dos especies deStenocereus: S. aragonii y.S. eichlamii, con base en dos caracteres de la semilla (paredes periclinales planas y microrrelieve finamente rugoso). El géneroPachycereus no es monofilético de acuerdo con la clasificación más reciente;Pachycereus hollianus yP. lepidanthus carecen de zona fértil diferenciada y representan el grupo hermano de las restantes especies de la subtribu Pachycereinae;P. fulviceps está relacionado con los génerosCephalocereus yNeobuxbaumia por el tipo de zona nectarial, el color de la pulpa del fruto y los cristales prismáticos de oxalato de calcio en células epidérmicas;P. grandis, P. pecten-aboriginum, P. pringlei, P. tepamo yP. weberi se resuelven como grupo monofilético por presentar un surco angosto y una flor con constricción en la base del tubo. Este grupo de cinco especies debe representar al géneroPachycereus sensu stricto.