Secondary Metabolites and Biological Properties of Gesneriaceae Species

The family Gesneriaceae comprises ca. 150 genera and 3000 species, distributed in the tropics around the world. It is constituted of herbs, lianas, or shrubs, frequently with ornamental potential, due to the beauty of their flowers. Some species have been used in traditional medicine, mainly against fever, cough, colds, snakebite, pains, and infectious and inflammatory diseases. Although Gesneriaceae are a large family, only few species were chemically investigated, and this took place mainly in the last decade. In the present work, chemical and pharmacological studies on Gesneriaceae are reviewed based on original articles published. Altogether 300 compounds have been reported in Gesneriaceae species, including flavonoids, terpenes and steroids, phenolic glucosides, simple phenolics, quinones, lignans, xanthones, and compounds with unusual skeletons. Several species had been used in folk medicine, and some constituents have shown biological activities, such as antimicrobial, anti‐inflamatory, antioxidant, and antitumor properties.     

Flowering,fruiting and seed germination in Chilean rain forest myrtaceae: ecological and phylogenetic constraints

Phenological studies in plant communities have generally focused on taxonomically heterogeneous species assemblages, and have only occasionally examined the evolutionary and ecological constraints on the phenological patterns of species within a single family or a genus. Here, we determine relative importance of phylogenetic versus other constraints on the flowering and fruiting periods of 12 species and the germination ecology of 10 species of Myrtaceae sympatric to the temperate rainforest of Isla Grande de Chiloé (42 °Cs 30 S), in southern Chile.We found that, for most species in the family Myrtaceae, flowering was strongly aggregated in January and February. Although this pattern is consistent with the expectation of the 'facilitation' hypothesis (i.e., interspecific overlaps are maximized to attract pollinators), available evidence suggests that pollinators, mainly hymenopterans and dipterans, do not limit fruit production in these species of Myrtaceae in the temperate rainforest. In contrast to flowering, fruiting occurred all-year-round, showing greater segregation in time among the species. According to the their temporal patterns of fruit ripening, two functional groups were defined within the Myrtaceae: those that ripen their fruits immediately after flowering (species in the subtribe Myrtinae) and those in which green fruit develops slowly for several months before ripening (subtribe Myrciinae). Seed germination in the field occurred mainly between August and October. Lab assays showed that the species of Myrtaceae, subtribe Myrtinae, exhibited a long seed dormancy (>40 days), while the seeds of species in the subtribe Myrciinae often germinated within one week after harvesting. The analysis of the phenology of reproductive events in the species of Myrtaceae in this rainforest suggests that: (1) flowering periods patterns are constrained mainly by phylogenetic inertia at the family level, and (2) differences in fruiting patterns and dormancy periods are determined mainly by fruit and seed size, which in turn are associated primarily with phylogenetic closeness within the family, and secondarily with the activity of vertebrate seed dispersers.     

Cholestane-3beta,5alpha,6beta-triol inhibits osteoblastic differentiation and promotes apoptosis of rat bone marrow stromal cells

Converging lines of evidence suggest that oxidized lipids, long recognized as a risk factor in atherogenesis, also contribute to osteoporosis, but the underlying mechanism is not understood in detail. The effect of atherogenesis related factors including oxysterols on the differentiation and survival of marrow stromal cells (MSCs) would be very important in understanding the link between atherosclerosis and osteoporosis. In the present study, the effect of oxysterol cholestane-3beta,5alpha,6beta-triol (Triol) on osteoblastic differentiation and apoptosis of primary rat bone MSCs as well as the related mechanisms were studied. Triol inhibited MSCs osteoblastic differentiation as demonstrated by inhibition of alkaline phosphatase activity, osteocalcin secretion, and matrix mineralization. In the other aspect, Triol promoted MSCs apoptosis, as characterized by condensed or fragmented nuclei as well as active externalization of phosphatidyl serine to the cell surface. In addition, Triol was found to induce increases of intracellular Ca2+ and Ca2+-dependent reactive oxygen species generation in MSCs. These effects were involved in the action of Triol on apoptosis, but not on osteoblastic differentiation of MSCs. These results suggested that Triol might contribute to the decreased bone formation by inhibition of osteoblastic differentiation and promotion of apoptosis of MSCs, providing insights about common factors underlying the pathogenesis of atherosclerosis and osteoporosis.     

Chromatographic analysis of carotenol fatty acid esters in Physalis alkekengi and Hippophae rhamnoides

The carotenol fatty acid esters of two potentially valuable sources of plant carotenoids, sepals of Physalis alkekengi (Chinese lantern) and fruits of Hippophae rhamnoides (sea buckthorn), were separated by column chromatography and identified by HPLC-DAD and HPLC-MS. A chemical and an enzymatic hydrolysis were employed to identify the parent carotenoids and to remove the lipid components. Zeaxanthin and beta-cryptoxanthin esters represented the main fraction in P. alkekengi sepals and an important one in H. rhamnoides fruits. Beta-Cryptoxanthin palmitate and zeaxanthin dipalmitate were identified as major compounds in both plants. In P. alkekengi, the carotenoids were mainly (> 90%) esterified with palmitic acid, and a high proportion (> 80%) of saturated medium chain fatty acids was found (by GC-MS) in the total lipid extract. Although the total lipid extract of H. rhamnoides contained significant amounts of unsaturated fatty acids, especially oleic and palmitoleic acids, the xanthophylls were mainly esterified with saturated fatty acids. The oleoresins of both species represent potential sources of carotenoid esters and can be used as food additives, cosmetic ingredients or nutraceuticals.     

Hyperdominance in fruit production in the Brazilian Atlantic rain forest: the functional role of plants in sustaining frugivores

The availability of fruits is critical for tropical forests, where the majority of plant species rely upon animal vectors for seed dispersal. However, we do not know how fruit production is temporally distributed over species and families. Two plant families are particularly important in floristic inventories of Atlantic rain forests: Arecaceae, a few species of which are highly abundant; and Myrtaceae, which is abundant and displays outstanding species diversity. In this context, we asked whether hyperdominance occurs in fruit production in the Atlantic rain forest, and whether it occurs in the abundant species of Arecaceae and Myrtaceae. We investigated whether the temporal fruit production patterns differ between Myrtaceae, Arecaceae, and the plant community as a whole. We also applied a functional dispersion index to assess the temporal fruit diversity over a 2‐yr period, with regard to morphological and phenological traits. We found that the phenomenon of hyperdominance occurs in fruit production: five species accounted for more than half of the pulp biomass. Arecaceae fruit biomass peaked at the end of wet season, overlapping with the community peak; whereas Myrtaceae species fruited throughout the year and were an important resource during periods of food scarcity. Myrtaceae filled more of the fruit morphospace over time because their fruits exhibit a large range of morphologies and phenological strategies. Our results demonstrated the importance of combining phenology and fruit morphology in the evaluation of resource availability, which revealed periods of high fruit diversity that could support a range of frugivore sizes and maintain overall ecosystem functionality.     

Analysis of the differences in the folding kinetics of structurally homologous proteins based on predictions of the gross features of residue contacts

It is a general notion that proteins with very similar three-dimensional structures would show very similar folding kinetics. However, recent studies reveal that the folding kinetic properties of some proteins contradict this thought (i.e., the members in a same protein family fold through different pathways). For example, it has been reported that some beta-proteins in the intracellular lipid-binding protein family fold through quite different pathways (Burns et al., Proteins 1998;33:107-118). Similar differences in folding kinetics are also observed in the members of the globin family (Nishimura et al., Nat Struct Biol 2000;7:679-686). In our study, we examine the possibility of predicting qualitative differences in folding kinetics of the intracellular lipid-binding proteins and two globin proteins (i.e., myoglobin and leghemoglobin). The problem is tackled by means of a contact map based on the average distance statistics between residues, the Average Distance Map (ADM), as constructed from sequence. The ADMs for the three proteins show overall similarity, but some local differences among maps are also observed. Our results demonstrate that some properties of the protein folding kinetics are consistent with local differences in the ADMs. We also discuss the general possibility of predicting folding kinetics from sequence information.     

Locus-specific protocol for nine different innate immune genes (antimicrobial peptides: β-defensins) across passerine bird species reveals within-species coding variation and a case of trans-species polymorphisms

We present a locus-specific protocol suitable for the investigation, from extracted DNA, into natural inter- and intra-specific genetic variation in a group of nine innate immune genes, all belonging to the β-defensin gene family. The products of these genes encode peptides with antimicrobial properties and form part of the innate immune system. The protocol amplifies the exon coding for the peptide that interacts with invading pathogens and is applicable across a wide range of passerine bird species, although with varying success depending on species. In several species tested, we found individuals to be heterozygous at several of the genes, highlighting the existence of coding genetic variation in this part of the immune system. Furthermore, for several of the genes, alleles have been conserved at the amino acid level across taxonomically distant bird species. In one case, we observed the existence of trans-species polymorphisms, often taken as evidence of balancing selection. The method will make it possible to investigate a part of the immune system for which there exists very little information of the genetic variation in wild vertebrate populations, thus making it possible to start investigating the selective forces under which the genes are evolving and the extent to which the found genetic variation is associated with pathogen susceptibility in wild populations.     

Gallery forest types and their environmental correlates in a Colombian savanna landscape

Savanna regions may contain significant areas of forest, usually in riparian environments. We described forest composition and quantified environmental variables for 80 plots in the watershed of the Yucao River (2550 km²), a tributary to the Meta River (Orinoco basin) in the eastern plains of Colombia. The total sampled area of 3.2 ha contained 147 tree species (110 genera and 45 families) with diameter at breast height >5 cm. The families represented by most species were Leguminosae (15 species), Arecaceae (11 species), Rubiaceae (9), Chrysobalanaceae (7), Euphorbiaceae (7), Melastomataceae (7) and Myrtaceae (7). Although most of the tree species have wide distribution ranges in humid neotropical forests, and few are endemic to the region, comparison with other neotropical forests suggests that the combination of taxa is characteristic for neotropical gallery forests. Cluster analysis allowed the definition of five forest types. They show important differences in a number of environmental variables, especially those related to inundation regimes and soil properties. Canonical correspondence analysis indicates good correlation of the first ordination axis with the size of the stream with which the forest is associated, the height to which the water rises during floods and variables related to soil fertility. The second ordination axis differentiates between soils with high or low water content in the dry season, high or low organic matter content and low or high bulk density. The results demonstrate the role of spatial variability and interactions of floodplain morphology, hydrological regime, and soil properties in structuring this important riparian community.     

14-3-3ε marks the amyloid-stimulated microglia long-term activation

Microglia-mediated inflammation in the central nervous system is a hallmark of the pathogenesis of several neurodegenerative diseases including Alzheimer's disease. Microglial cells activation follows the deposition of amyloid β fibrils and it is generally considered a triggering factor in the early steps of the onset of Alzheimer's disease. Although the initial engagement of microglia seems to play a neuroprotective role, many lines of evidence indicate that a persistent activation with the production of proinflammatory molecules contributes to dismantle neuronal activity and to induce neuronal loss occurring in neurodegenerative diseases. To date, limited proteomic data are available on activated microglial cells in response to extracellular amyloidogenic peptides. In this study, murine microglial cells have been employed to investigate the effects of amyloid β peptides in triggering microglial activation. The response was monitored at the proteome level through a two-dimensional gel electrophoresis-based approach. Results show only a limited number of differentially expressed proteins, among these a more acidic species of the cytosolic actin, and the 14-3-3ε protein, found significantly upregulated in Aβ-activated cells. 14-3-3ε belongs to a regulatory protein family involved in important cellular processes, including those leading to neurodegenerative diseases, and thus its increased expression suggests a role of this protein in tuning microglia activation.     

Role of endogenous TGF-β family in myogenic differentiation of C2C12 cells

The present study evaluated endogenous activities and the role of BMP and transforming growth factor-β (TGF-β), representative members of the TGF-β family, during myotube differentiation in C2C12 cells. Smad phosphorylation at the C-terminal serines was monitored, since TGF-β family members signal via the phosphorylation of Smads in a ligand-dependent manner. Expression of phosphorylated Smad1/5/8, which is an indicator of BMP activity, was higher before differentiation, and rapidly decreased after differentiation stimulation. Differentiation-related changes were consistent with those in the expression of Ids, well-known BMP-responsive genes. Treatment with inhibitors of BMP type I receptors or noggin in C2C12 myoblasts down-regulated the expression of myogenic regulatory factors, such as Myf5 and MyoD, leading to impaired myotube formation. Addition of BMP-2 during the myoblast phase also inhibited myotube differentiation through the down-regulation of Myf5 and MyoD. In contrast to endogenous BMP activity, the phosphorylation of Smad2, a TGF-β-responsive Smad, was higher 8-16 days after differentiation stimulation. A-83-01, an inhibitor of TGF-β type I receptor, increased the expression of Myf5 and MyoD, and enhanced myotube formation. The present results reveal that endogenous activities of the TGF-β family are changed during myogenesis in a pathway-specific manner, and that the activities are required for myogenesis.     

Minimal surface as a model of beta-sheets

The purpose of this article is to present arguments based on experimental data that the beta-sheet structures in proteins are the result of the tendency to minimize surface areas. Thus, we propose the model that all beta-sheet structures are almost minimal surfaces, namely, their mean curvatures are nearly zero. To support this model, we chose 1740 disjoint beta-sheets with less than 10 strands from the all beta-protein class in a nonredundant 40% Structural Classification of Proteins (SCOP) database and applied the least-squares method to fit the minimal surface catenoid (and in some rare cases, the plane) to the beta-sheet structures. The fitting errors were extremely small: The error of 1729 beta-sheets with catenoid minimal surface is 0.90 +/- 0.55 A and the error of the remaining 11 flat sheets with the plane is 0.64 +/- 0.46 A. The fact that the commonly used models for some beta-sheet surfaces (i.e., the hyperboloid and strophoid) have very small mean curvatures (< 0.05) supports our model. Moreover, we showed that this model also includes the isotropically stressed configuration model proposed by Salemme, in which the intrastrand tendency of the individual chains to twist or coil is in equilibrium with the tendency of the interstrand hydrogen bonding to resist twisting of the sheet as a whole. As an application we used our model to quantify the two principal independent modes in the flexibility of beta-sheets, that is, the bending parameter of beta-sheets and the inclined angle of beta-strands in a sheet.     

Curitiba (Myrtaceae): A new genus from the Planalto of southern Brazil

In the course of monographic work in Myrtaceae it has become evident thatEugenia prismatica cannot be accommodated within any existing genus in the family. Therefore, a new genus,Curitiba, is described to contain it and the new combinationCuritiba prismatica is made.Curitiba is diagnosed by a combination of 4-angled hypanthia and fruits, ovules radiating from a central protruding placenta, and seed coats with wavy rows of papillae. A key is provided to distinguishCuritiba from the Caribbean and Mesoamerican genusMosiera, where the species also had been placed before this transfer, and from other 4-merous genera in Brazil of subtribe Myrtinae. In the Atlantic Forest of Brazil where the family Myrtaceae is one of the most species rich and ecologically important plant families, the discovery ofCuritiba reveals a unique evolutionary lineage, highlights the endemism present in the Atlantic Forest, and underscores, the urgent need for conservation of this rapidly disappearing and highly endangered biome.     

A Global overview of canids used in traditional medicines

Canids are among the mammal species most frequently used in traditional folk medicine around the world. In this context, this paper assesses the global use of canids in traditional folk medicine and their implications. Our review indicated that 19 species of canid are used in traditional medicine worldwide, representing 54.2% of described canid species. Of the species in medicinal use, two are listed as Endangered and three as near threatened on the IUCN Red List. For some species medicinal use represents an additional direct pressure that may have contributed to declines of natural populations. In addition, use of medicinal animals may have indirect impacts on the conservation of other species through the spread of disease. To minimize both harvest impacts and disease spread, guidance on use of medicinal species may be useful. This could include an exploration of the use of alternatives and implementation of sanitary measures.     

Resource Partitioning between Highly Eusocial Bees and Possible Impact of the Introduced Africanized Honey Bee on Native Stingless Bees in the Brazilian Atlantic Rainforest

The highly eusocial bee community of the neotropical Atlantic Rainforest was studied at Boracéia Biological Station in the state of São Paulo, Brazil. In this reserve, 17 species of stingless bees and the introduced Africanized honey bee were found, the latter being the most abundant flower visitor. Of all flowering plants, Asteraceae and Myrtaceae were particularly important as resources for bees. Trophic niche overlap between the various species of stingless bees is evident, and it was generally larger within the tribes Meliponini and Trigonini than between members of different tribes. Nevertheless, in the stingless bee community the competitive pressure is rather uniformly spread. The trophic niche of the Africanized honey bee can be positioned between those of Meliponini and Trigonini. Today this introduced species represents the main competitor in this bee community. However, its impact on native stingless bee populations is apparently buffered by mass-flowering trees which are the most important food plants of the indigenous highly eusocial bees.     

Computational analysis of glycoside hydrolase family 1 specificities

Glycoside hydrolase family 1 consists of beta-glucosidases, beta-galactosidases, 6-phospho-beta-galactosidases, myrosinases, and other enzymes having similar primary and tertiary structures but diverse specificities. Among these enzymes, beta-glucosidases hydrolyze cellobiose to glucose, and therefore they are key players in any cellulose to glucose process. All family members attack beta-glycosidic bonds between a pyranosyl glycon and an aglycon, but most have little specificity for the aglycon or for the bond configuration. Furthermore, glycon specificity is not absolute. Sixteen family members (six beta-glucosidases, two cyanogenic beta-glucosidases, one 6-phospho-beta-galactosidase, two myrosinases, and five beta-glycosidases) have known tertiary structures. We have used automated docking to computationally bind disaccharides with allopyranosyl, galactopyranosyl, glucopyranosyl, mannopyranosyl, 6-phosphogalactopyranosyl, and 6-phosphoglucopyranosyl glycons, all linked by beta-(1,2), beta-(1,3), beta-(1,4), and beta-(1,6)-glycosidic bonds to beta-glucopyranoside aglycons, along with beta-(1,1-thio)-allopyranosyl, -galactopyranosyl, -glucopyranosyl, and -mannopyranosyl) beta-glucopyranosides, into all of these structures to investigate the structural determinants of their enzyme specificities. The following are the eight active-site residues: Glu191, Thr194, Phe205, Asn285, Arg336, Asn376, Trp378, and Trp465 (Zea mays beta-glucosidase numbering), that control a significant amount of glycon specificity. (c) 2008 Wiley Periodicals, Inc. Biopolymers 89: 1021-1031, 2008.This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com.     

Convergence in tent architecture and tent-making behavior among neotropical and paleotropical bats

Fifteen species of neotropical and three species of paleotropical bats are known either to roost in or to make tents in over 80 species of vascular plants. We summarize the current knowledge of bat-tent architecture, report two new styles of tents (conical and apical) from the Paleotropics, compare the similarity in tents constructed, or used, by neotropical and paleotropical bats, and consider possible functions of tents. Seven styles of tents are known from the Neotropics, three (conical, palmate umbrella, and apical tents) are known from both the Neo- and the Paleotropics, and one (stem tent) is unique to the Paleotropics. In the Neotropics tent-roosting and/or tent-making appears to be a behavior unique to the diverse microchiropteran family Phyllostomidae (subfamily Phyllostomatinae: tribe Stenodermatini), and in the Paleotropics two members of the megachiropteran family Pteropodidae and one member of the microchiropteran family Vespertilionidae are known to construct or roost in tents. Despite the variety of plant taxa used by bats in tent construction, there appears to be a limited number of different leaf forms that can be altered by bats and used as tents. We suggest that the similarity in tent architecture observed among the neotropical and paleotropical bats is a consequence of convergence in leaf morphology among forest understory plants. The congruence in tent-making/roosting behavior observed in members of the Stenodermatini and the Pteropodidae (genusCynopterus) suggests a phylogenetic influence on these behaviors. The similarity in tent-making and/or tent-roosting behavior and life-history traits (small, <70 g, mostly foliage-roosting frugivores) among these divergent neotropical and paleotropical taxa supports a convergence hypothesis in which members of these groups have become ecological equivalents. Although actual tent-making has been observed in only one bat species to date, we suggest that the principal selective force leading to the evolution of tent-making is a polygynous mating system whereby males construct tents to gain access to females. Tents in turn provide resources that offer protection from predators and inclement weather.     

Thermoregulatory ability and mechanism do not differ consistently between neotropical and temperate butterflies

Climate change is a major threat to species worldwide, yet it remains uncertain whether tropical or temperate species are more vulnerable to changing temperatures. To further our understanding of this, we used a standardised field protocol to (1) study the buffering ability (ability to regulate body temperature relative to surrounding air temperature) of neotropical (Panama) and temperate (the United Kingdom, Czech Republic and Austria) butterflies at the assemblage and family level, (2) determine if any differences in buffering ability were driven by morphological characteristics and (3) used ecologically relevant temperature measurements to investigate how butterflies use microclimates and behaviour to thermoregulate. We hypothesised that temperate butterflies would be better at buffering than neotropical butterflies as temperate species naturally experience a wider range of temperatures than their tropical counterparts. Contrary to our hypothesis, at the assemblage level, neotropical species (especially Nymphalidae) were better at buffering than temperate species, driven primarily by neotropical individuals cooling themselves more at higher air temperatures. Morphology was the main driver of differences in buffering ability between neotropical and temperate species as opposed to the thermal environment butterflies experienced. Temperate butterflies used postural thermoregulation to raise their body temperature more than neotropical butterflies, probably as an adaptation to temperate climates, but the selection of microclimates did not differ between regions. Our findings demonstrate that butterfly species have unique thermoregulatory strategies driven by behaviour and morphology, and that neotropical species are not likely to be more inherently vulnerable to warming than temperate species.     

Galactomannanases Man2 and Man5 from Thermotoga species: growth physiology on galactomannans, gene sequence analysis, and biochemical properties of recombinant enzymes

The enzymatic hydrolysis of mannan-based hemicelluloses is technologically important for applications ranging from pulp and paper processing to food processing to gas and oil well stimulation. In many cases, thermostability and activity at elevated temperatures can be advantageous. To this end, the genes encoding beta-mannosidase (man2) and beta-mannanase (man5) from the hyperthermophilic bacteria Thermotoga neapolitana 5068 and Thermotoga maritima were isolated, cloned, and expressed in Escherichia coli. The amino acid sequences for the mannosidases from these organisms were 77% identical and corresponded to proteins with an M(r) of approximately 92 kDa. The translated nucleotide sequences for the beta-mannanase genes (man5) encoded polypeptides with an M(r) of 76 kDa that exhibited 84% amino acid sequence identity. The recombinant versions of Man2 and Man5 had similar respective biochemical and biophysical properties, which were also comparable to those determined for the native versions of these enzymes in T. neapolitana. The optimal temperature and pH for the recombinant Man2 and Man5 from both organisms were approximately 90 degrees C and 7.0, respectively. The presence of Man2 and Man5 in these two Thermotoga species indicates that galactomannan is a potential growth substrate. This was supported by the fact that beta-mannanase and beta-mannosidase activities were significantly stimulated when T. neapolitana was grown on guar or carob galactomannan. Maximum cell densities increased by at least tenfold when either guar or carob galactomannan was added to the growth medium. For T. neapolitana grown on guar at 83 degrees C, Man5 was secreted into the culture media, whereas Man2 was intracellular. These localizations were consistent with the presence and lack of signal peptides for Man5 and Man2, respectively. The identification of the galactomannan-degrading enzymes in these Thermotoga species adds to the list of biotechnologically important hemicellulases produced by members of this hyperthermophilic genera.     

Evolution of exceptional species richness among lineages of fleshy-fruited Myrtaceae

MethodsBayesian phylogenetic analyses of plastid and nuclear DNA sequences were used to estimate intertribal relationships and lineage divergence times in Myrtaceae. Focusing on the fleshy-fruited tribes, a variety of statistical approaches were used to assess diversification rates and diversification rate shifts across the family.ConclusionsFleshy fruits have evolved independently in Syzygieae and Myrteae, and this is accompanied by exceptional diversification rate shifts in both instances, suggesting that the evolution of fleshy fruits is a key innovation for rainforest Myrtaceae. Noting the scale dependency of this hypothesis, more complex explanations may be required to explain diversification rate shifts occurring within the fleshy-fruited tribes, and the suggested phylogenetic hypothesis provides an appropriate framework for this undertaking.