Evaluation in Cameroon of a Novel,Simplified Methodology to Assist Molecular Microbiological Analysis of V. cholerae in Resource-Limited Settings

BackgroundVibrio cholerae is endemic in South Asia and Africa where outbreaks of cholera occur widely and are particularly associated with poverty and poor sanitation. Knowledge of the genetic diversity of toxigenic V. cholerae isolates, particularly in Africa, remains scarce. The constraints in improving this understanding is not only the lack of regular cholera disease surveillance, but also the lack of laboratory capabilities in endemic countries to preserve, store and ship isolates in a timely manner. We evaluated the use of simplified sample preservation methods for molecular characterization using multi-locus variable-number tandem-repeat analysis (MLVA) for differentiation of Vibrio cholerae genotypes.ConclusionsCollecting V. cholerae using simplified laboratory methods in remote and low-resource settings allows for subsequent advanced molecular characterization of V. cholerae O1. These simplified DNA preservation methods identify V. cholerae and make possible timely information regarding the genetic diversity of V. cholerae; our results set the stage for continued molecular epidemiological research to better understand the transmission and dissemination of V. cholerae in Africa and elsewhere worldwide.     

Gels prepared from egg yolk and its fractions for tissue engineering

New biomaterials prepared from egg yolk and its main fractions (plasma and granules) have been developed for use in tissue engineering. Protein gels obtained via transglutaminase cross‐linking were characterized by rheometry, texturometry and scanning electron microscopy. All the gels exhibited suitable physical and mechanical characteristics for use as potential biomaterials in skin regeneration. Specifically, results showed that these materials presented a compact, uniform structure, with granular gel being found to be the most resistant as well as the most elastic material. Accordingly, these gels were subsequently evaluated as scaffolds for murine fibroblast growth. The best results were obtained with granule gels. Not only adhesion and cell growth were detected when using these gels, but also continuous coatings of cells growing on their surface. These findings can be attributed to the higher protein content of this fraction and to the particular structure of its proteins. Thus, granules have proved to be an interesting potential raw material for scaffold development. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1577–1583, 2016     

High-performance targeted mass spectrometry with precision data-independent acquisition reveals site-specific glycosylation macroheterogeneity

Protein glycosylation is a critical post-translational modification that regulates the structure, stability, and function of many proteins. Mass spectrometry is currently the preferred method for qualitative and quantitative characterization of glycosylation. However, the inherent heterogeneity of glycosylation makes its analysis difficult. Quantification of glycosylation occupancy, or macroheterogeneity, has proven to be especially challenging. Here, we used a variation of high-resolution multiple reaction monitoring (MRM^HR) or pseudo-MRM for targeted data-independent acquisition that we term SWAT (sequential window acquisition of targeted fragment ions). We compared the analytical performance of SWATH (sequential window acquisition of all theoretical fragment ions), SWAT, and SRM (selected reaction monitoring) using a suite of synthetic peptides spiked at various concentrations into a complex yeast tryptic digest sample. SWAT provided superior analytical performance to SWATH in a targeted approach. We then used SWAT to measure site-specific N-glycosylation occupancy in cell wall glycoproteins from yeast with defects in the glycosylation biosynthetic machinery. SWAT provided robust measurement of occupancy at more N-glycosylation sites and with higher precision than SWATH, allowing identification of novel glycosylation sites dependent on the Ost3p and Ost6p regulatory subunits of oligosaccharyltransferase.     

Vocal Cues to Identity: Pied Babblers Produce Individually Distinct But Not Stable Loud Calls

The ability to identify social partners can play a key role in the coordination of social behaviours in group‐living animals. Coordinating social behaviours over long distances becomes problematic, as cues to identity are often limited to one or two sensory modalities. This limitation can often select for strong individuality in those cues used for long‐distance communication. Pied babblers, Turdoides bicolor, produce a number of different types of ‘loud calls’ which are frequently used to signal to individuals beyond the range of visual or olfactory pathways of communication. Here, we show that three of these ‘loud call’ types, the v‐shaped chatter, the double note ascending chatter and the atonal chatter, are each individually distinct. We hypothesise that individuality in the three loud call types tested here may represent a possible pathway to social recognition in this species that may have important consequences for social interactions. However, we also found that the atonal chatter was unstable between years suggesting that this particular call type may not be a reliable long‐term indicator to identity which may affect long‐term recognition in this species.     

Variability on microevolutionary and macroevolutionary scales: a review on patterns of morphological variation in Cnidaria Medusozoa

Members of Cnidaria Medusozoa are known for their wide morphological variation, which is expressed on many different levels, especially in different phases of the life cycle. Difficulties in interpreting morphological variations have posed many taxonomic problems, since intraspecific morphological variations are often misinterpreted as interspecific variations and vice-versa, hampering species delimitation. This study reviews the patterns of morphological variation in the Medusozoa, to evaluate how different interpretations of the levels of variation may influence the understanding of the patterns of diversification in the group. Additionally, we provide an estimate of the cryptic diversity in the Hydrozoa, based on COI sequences deposited in GenBank. Morphological variations frequently overlap between microevolutionary and macroevolutionary scales, contributing to misinterpretations of the different levels of variation. In addition, most of the cryptic diversity described so far for the Medusozoa is a result of previously overlooked morphological differences, and there is still great potential for discovering cryptic lineages in the Hydrozoa. We provide evidence that the number of species in the Medusozoa is misestimated and emphasize the necessity of examining different levels of morphological variations when studying species boundaries, in order to avoid generalizations and misinterpretations of morphological characters.