The NLRP12 pyrin domain: structure, dynamics, and functional insights

The initial line of defense against infection is sustained by the innate immune system. Together, membrane-bound Toll-like receptors and cytosolic nucleotide-binding domain and leucine-rich repeat-containing receptors (NLR) play key roles in the innate immune response by detecting bacterial and viral invaders as well as endogenous stress signals. NLRs are multi-domain proteins with varying N-terminal effector domains that are responsible for regulating downstream signaling events. Here, we report the structure and dynamics of the N-terminal pyrin domain of NLRP12 (NLRP12 PYD) determined using NMR spectroscopy. NLRP12 is a non-inflammasome NLR that has been implicated in the regulation of Toll-like receptor-dependent nuclear factor-κB activation. NLRP12 PYD adopts a typical six-helical bundle death domain fold. By direct comparison with other PYD structures, we identified hydrophobic residues that are essential for the stable fold of the NLRP PYD family. In addition, we report the first in vitro confirmed non-homotypic PYD interaction between NLRP12 PYD and the pro-apoptotic protein Fas-associated factor 1 (FAF-1), which links the innate immune system to apoptotic signaling. Interestingly, all residues that participate in this protein:protein interaction are confined to the α2-α3 surface, a region of NLRP12 PYD that differs most between currently reported NLRP PYD structures. Finally, we experimentally highlight a significant role for tryptophan 45 in the interaction between NLRP12 PYD and the FAF-1 UBA domain.     

Directed evolution of colE1 plasmid replication compatibility: a fast tractable tunable model for investigating biological orthogonality

Plasmids of the ColE1 family are among the most frequently used in molecular biology. They were adopted early for many biotechnology applications, and as models to study plasmid biology. Their mechanism of replication is well understood, involving specific interactions between a plasmid encoded sense-antisense gene pair (RNAI and RNAII). Due to such mechanism, two plasmids with the same origin cannot be stably maintained in cells—a process known as incompatibility. While mutations in RNAI and RNAII can make colE1 more compatible, there has been no systematic effort to engineer new compatible colE1 origins, which could bypass technical design constraints for multi-plasmid applications. Here, we show that by diversifying loop regions in RNAI (and RNAII), it is possible to select new viable colE1 origins compatible with the wild-type one. We demonstrate that sequence divergence is not sufficient to enable compatibility and pairwise interactions are not an accurate guide for higher order interactions. We identify potential principles to engineer plasmid copy number independently from other regulatory strategies and we propose plasmid compatibility as a tractable model to study biological orthogonality.     

Isolation and characterization of microsatellite loci in Epidendrum puniceoluteum, an endemic orchid from the Atlantic Rainforest

Epidendrum puniceoluteum is an endemic orchid of Atlantic Rainforest, restricted to few populations only due to the destruction and fragmentation of its native habitat. Here, we report on the development of 10 microsatellite markers isolated from this orchid species. Genetic variability was characterized in two distant populations from Brazil coast. The number of alleles observed for each locus ranged from two to 12 and with an average of 6.4 alleles per locus. These microsatellites should be valuable tools for studying both fine-scale genetic structure of scattered E. puniceoluteum population and patterns will be useful genetic markers for other closely related taxa.     

Downregulation of metallothionein 2A reduces migration,invasion and proliferation activities in human squamous cell carcinoma cells

Molecular Biology Reports - The invasive behaviour of squamous cell carcinoma (SCC), a common malignant tumour of the mouth, is a process mediated by cell proliferation, extracellular matrix...     

Effects of age and mannanoligosaccharides supplementation on production of volatile fatty acids in the caecum of rabbits

This trial studied the effect of including mannanoligosaccharides (MOS, Bio-Mos^®, Alltech Inc., USA) in the diet on the caecal volatile fatty acids (VFA) and pH of rabbits from 34 to 90 days of age. Three experimental diets were compared: control diet, zinc bacitracin (ZnBac) diet (control diet with 0.1 g ZnBac/kg feed) and MOS diet (control diet with 2.0 g MOS/kg feed). Rabbits were slaughtered at 34, 48, 69 and 90 days of age and caecal contents were collected and analyzed for dry matter, pH and VFA concentration. The empty caecum and caecal contents weights relative to live weight were also determined.Age affected (P<0.0001) VFA concentration and pH values in the caecum. The pH decreased with age whereas VFA concentration increased. Rabbits fed MOS had higher (P<0.05) VFA and tended (P=0.098) to had lower pH in the caecum than rabbits fed ZnBac and control diets. Acetic, propionic and butyric acids concentrations in the caecum increased with MOS but its molar proportions were similar among diets. Diet had no effect on empty caecum weight and caecal contents weight and dry matter concentration. There was no interaction effect between diet and age. From 34 to 90 days of age, VFA production was higher (P<0.05) in the caecum of rabbits fed MOS than in those fed ZnBac diet and control diet.The addition of MOS to the diet increased the VFA concentration in the caecum of growing rabbits from 34 to 90 days of age.     

Vocal Signature in Wild Infant Chimpanzees

A large array of communication signals supports the fission/fusion social organization in chimpanzees, and among them the acoustic channel plays a large part because of their forest habitat. Adult vocalizations convey social and ecological information to their recipients allowing them to obtain cues about an ongoing event from calls only. In contrast to adult vocalizations, information encoded in infant calls had been hardly investigated. Studies mainly focused on vocal development. The present article aims at assessing the acoustic cues that support individual identity coding in infant chimpanzees. By analyzing recordings performed in the wild from seven 3‐year‐old infant chimpanzees, we showed that their calls support a well‐defined individual vocal signature relying on spectral cues. To assess the reliability of the signature across the calls of an individual, we defined two subsets of recordings on the basis of the characteristics of the frequency modulation (whimpers and screams) and showed that both call types present a reliable vocal signature. Early vocal signature may allow the mother and other individuals in the group to identify the infant caller when visual contact is broken. Chimpanzee mothers may have developed abilities to cope with changing vocal signatures while their infant, still vulnerable, gains in independence in close habitat. Am. J. Primatol. 75:324‐332, 2013. © 2012 Wiley Periodicals, Inc.     

Antioxidant,anti-inflammatory and healing potential of ethyl acetate fraction of Bauhinia ungulata L. (Fabaceae) on in vitro and in vivo wound model

The present work aimed to investigate the antioxidant, anti-inflammatory and wound healing potential of ethyl acetate fraction from Bauhinia ungulata L. (FABU) on in vitro and in vivo models. Wound healing assay using human lung adenocarcinoma A549 cell line was employed to evaluate the ability of FABU in modulating cell migration. In addition, a surgical wound model in C57BL/6 mice was used to study the healing potential of FABU incorporated into gel carbomer 940 (Carbopol®). Evaluation of lipid peroxidation, inflammatory and anti-inflammatory mediator gene expression, rate of wound closure, and histological analysis were done. FABU significantly reduced the gap area in in vitro wound healing assay, 24 h after treatment. In the animal model, FABU at 0.5% topically applied once-daily for 5 days to the surgical wounds significantly reduced the lesion area. Moreover, it significantly decreased the levels of lipid peroxidation in the lesions and decreased the relative gene expression levels of IL-1β and TNF-α in the injured region. In conclusion, our study suggests that Bauhinia ungulata can effectively promote the wound healing, probably by regulating the inflammatory environment during the early stages of the process.

Graphic abstract

     

Evolution of territoriality in Hylinae treefrogs: Ecological and morphological correlates and lineage diversification

Given the diverse nature of traits involved in territorial defence, they may respond to different selective pressures and then exhibit distinct patterns of evolution. These selective pressures also may cause territorial behaviour to be associated with environmental and morphological variables. Such associations, however, have mostly been studied at the intraspecific level, being phylogenetic analyses of territoriality in a broad taxonomic framework rare in the literature. We used the anuran subfamily Hylinae to test (1) whether two territorial-behaviour traits with different levels of aggression—territorial call and physical combat—are evolutionarily more labile than a morphological trait used in physical combat—the spine-shaped prepollex; (2) whether reproduction in lentic waters and phytotelmata, as well as resource scarcity, might favour the occurrence of territoriality; (3) if physical combat is more important than territorial call for the evolution of body size and sexual size dimorphism and (4) the relationships between territorial-behaviour traits and lineage diversification. We mainly used the literature to build two datasets with different levels of certainty. Territorial-behaviour traits exhibited intermediate levels of phylogenetic signal in Hylinae, whereas the phylogenetic signal for the presence of the spine-shaped prepollex was strong. We found support for the hypothesis that reproduction in lentic water favours the occurrence of territorial behaviour, because the expression of territorial-behaviour traits was more associated with reproduction in lentic than in lotic waters. Territorial-behaviour traits were not correlated with annual precipitation nor with habitat complexity. Body size and sexual size dimorphism were not correlated with the presence of territorial call nor with physical combat. We identified negative correlations between diversification rates and physical combat. Relationships of territorial call and physical combat with diversification rates suggest that these territorial behaviours influence evolutionary processes in different ways.     

Histogenesis and histochemistry of the secretion plate: detection of glycans and neutral glycoproteins synthetized by the epithelial components of the gizzard mucosa of Gallus gallus

A study about the genesis of the inner lining which covers the mucous membrane of the chicken gizzard was made and histochemical test were employed to detect the presence of carbohydrate in the epithelial components of the mucous membrane. The histochemical treatment of the apical border of gizzard epithelium from the 5th, 7th, 9th, 12th, and 15th d of incubation revealed the absence of glycoprotein (with 1.2-glycol containing hexoses), sialic acid, and glycogen but weakly acidic sulphated mucosubstances were present. The same was observed in the medium 1/3 of the gizzard epithelium on 12th and 15th d. The upper 1/3 of the buddings, precursor of gizzard glands, on the 15th d, besides exhibiting the same histochemical characteristics of apical border, revealed, clearly visible, strongly sulphated mucosubstances. The histochemical reactions of the epithelium which covers the free surface and lines the pits from the 18th d of incubation and from the young chicken (0.5 h, 1 d, 3 d, 7 d, 3 weeks and 8 weeks) revealed the absence of glycoprotein (with 1.2-glycol containing hexoses), sialic acid, and glycogen, however, weakly acidic sulphated mucosubstances and strongly sulphated epithelial acidic mucin were present. On the 18th d, the glands presented a lumen and the accumulation of intraepithelial secretion displaced the upper part of the superficial epithelium, splitting, it.     

Effect of seven species of the family Asteraceae on longevity and nutrient levels of Episyrphus balteatus

Adult hoverflies feed on pollen, nectar and honeydews. Thus, the implementation of floristically diverse resources in agroecosystems can enhance biological control of pests by increasing populations of these natural enemies. The objective of this work was to study the effect of seven species of Asteraceae (Andryala integrifolia L., Anthemis arvensis L., Calendula arvensis L., Carduus tenuiflorus Curtis, Chamaemelum nobile (L.) All., Coleostephus myconis (L.) Rchb.f. and Crepis vesicaria L.) on adult longevity and nutrient levels of Episyrphus balteatus (De Geer) (Diptera: Syrphidae). Crepis vesicaria and C. nobile gave the highest mean longevities while, for the other plant species, longevities were not significantly different from those in the water only treatment. In addition, all species tested resulted in significantly lower longevities than the positive control (glucose). In terms of nutrient levels, C. nobile generated high levels of total sugars and C. vesicaria high levels of lipids which suggests the consumption of flower resources. However, the majority of the plants tested would not allow hoverflies to complete their life cycle. Thus, further experimentation needs to be done before considering these plants for implementation in agroecosystems.