Single-cell analysis identifies a key role for Hhip in murine coronal suture development.

Craniofacial development depends on formation and maintenance of sutures between bones of the skull. In sutures, growth occurs at osteogenic fronts along the edge of each bone, and suture mesenchyme separates adjacent bones. Here, we perform single-cell RNA-seq analysis of the embryonic, wild type murine coronal suture to define its population structure. Seven populations at E16.5 and nine at E18.5 comprise the suture mesenchyme, osteogenic cells, and associated populations. Expression of Hhip, an inhibitor of hedgehog signaling, marks a mesenchymal population distinct from those of other neurocranial sutures. Tracing of the neonatal Hhip-expressing population shows that descendant cells persist in the coronal suture and contribute to calvarial bone growth. In Hhip-/- coronal sutures at E18.5, the osteogenic fronts are closely apposed and the suture mesenchyme is depleted with increased hedgehog signaling compared to those of the wild type. Collectively, these data demonstrate that Hhip is required for normal coronal suture development.

Tapping into non-English-language science for the conservation of global biodiversity.

The widely held assumption that any important scientific information would be available in English underlies the underuse of non-English-language science across disciplines. However, non-English-language science is expected to bring unique and valuable scientific information, especially in disciplines where the evidence is patchy, and for emergent issues where synthesising available evidence is an urgent challenge. Yet such contribution of non-English-language science to scientific communities and the application of science is rarely quantified. Here, we show that non-English-language studies provide crucial evidence for informing global biodiversity conservation. By screening 419,679 peer-reviewed papers in 16 languages, we identified 1,234 non-English-language studies providing evidence on the effectiveness of biodiversity conservation interventions, compared to 4,412 English-language studies identified with the same criteria. Relevant non-English-language studies are being published at an increasing rate in 6 out of the 12 languages where there were a sufficient number of relevant studies. Incorporating non-English-language studies can expand the geographical coverage (i.e., the number of 2° × 2° grid cells with relevant studies) of English-language evidence by 12% to 25%, especially in biodiverse regions, and taxonomic coverage (i.e., the number of species covered by the relevant studies) by 5% to 32%, although they do tend to be based on less robust study designs. Our results show that synthesising non-English-language studies is key to overcoming the widespread lack of local, context-dependent evidence and facilitating evidence-based conservation globally. We urge wider disciplines to rigorously reassess the untapped potential of non-English-language science in informing decisions to address other global challenges. Please see the Supporting information files for Alternative Language Abstracts.

Simulated climate change, but not predation risk, accelerates Aedes aegypti emergence in a microcosm experiment in western Amazonia.

Climate change affects individual life-history characteristics and species interactions, including predator-prey interactions. While effects of warming on Aedes aegypti adults are well known, clarity the interactive effects of climate change (temperature and CO2 concentration) and predation risk on the larval stage remains unexplored. In this study, we performed a microcosm experiment simulating temperature and CO2 changes in Manaus, Amazonas, Brazil, for the year 2100. Simulated climate change scenarios (SCCS) were in accordance with the Fourth Assessment Report of Intergovernmental Panel on Climate Change (IPCC). Used SCCS were: Control (real-time current conditions in Manaus: average temperature is ~25.76°C ± 0.71°C and ~477.26 ± 9.38 parts per million by volume (ppmv) CO2); Light: increase of ~1,7°C and ~218 ppmv CO2; Intermediate: increase of ~2.4°C and ~446 ppmv CO2; and Extreme: increase of ~4.5°C and ~861 ppmv CO2, all increases were relative to a Control SCCS. Light, Intermediate and Extreme SCCS reproduced, respectively, the B1, A1B, and A2 climatic scenarios predicted by IPCC (2007). We analyzed Aedes aegypti larval survivorship and adult emergence pattern with a factorial design combining predation risk (control and predator presence-Toxorhynchites haemorrhoidalis larvae) and SCCS. Neither SCCS nor predation risk affected Aedes aegypti larval survivorship, but adult emergence pattern was affected by SCCS. Accordingly, our results did not indicate interactive effects of SCCS and predation risk on larval survivorship and emergence pattern of Aedes aegypti reared in SCCS in western Amazonia. Aedes aegypti is resistant to SCCS conditions tested, mainly due to high larval survivorship, even under Extreme SCCS, and warmer scenarios increase adult Aedes aegypti emergence. Considering that Aedes aegypti is a health problem in western Amazonia, an implication of our findings is that the use of predation cues as biocontrol strategies will not provide a viable means of controlling the accelerated adult emergence expected under the IPCC climatic scenarios.

Implementation of a Zebrafish Health Program in a Research Facility: A 4-Year Retrospective Study.

In the past two decades, zebrafish (Danio rerio)-based research has contributed to significant scientific advances. Still, husbandry and health programs did not evolve at the same pace, as evidenced by the absence of general guidelines. Health monitoring is essential to animal welfare, to permit animal exchanges across facilities, to contribute to robust experimental results, and for data reproducibility. In this study, we report a health program implemented in a zebrafish research facility to prevent, monitor, and control pathogen, and disease dissemination. This program includes quarantine, routine health screening of sentinels, and nonroutine screenings of retired animals and sick/moribund individuals. An extensive list of clinical signs, lesions, and pathogens was monitored based on: daily observation of fish, necropsy, histology, and bacterial culture. The results indicate that the combined analysis of sentinels with the evaluation of sick/moribund animals enables a comprehensive description not only of pathogen prevalence but also of clinical and histopathologic lesions of resident animals. The establishment of a quarantine program revealed to be effective in the reduction of Pseudoloma neurophilia frequency in the main aquaria room. Finally, characterization of the colony health status based on this multiapproach program shows a low prevalence of lesions and pathogens in the facility.

Noncoding RNAs of the Ultrabithorax domain of the Drosophila bithorax complex.

RNA transcripts without obvious coding potential are widespread in many creatures, including the fruit fly, Drosophila melanogaster. Several noncoding RNAs have been identified within the Drosophila bithorax complex. These first appear in blastoderm stage embryos, and their expression patterns indicate that they are transcribed only from active domains of the bithorax complex. It has been suggested that these noncoding RNAs have a role in establishing active domains, perhaps by setting the state of Polycomb Response Elements A comprehensive survey across the proximal half of the bithorax complex has now revealed nine distinct noncoding RNA transcripts, including four within the Ultrabithorax transcription unit. At the blastoderm stage, the noncoding transcripts collectively span ∼75% of the 135 kb surveyed. Recombination-mediated cassette exchange was used to invert the promoter of one of the noncoding RNAs, a 23-kb transcript from the bxd domain of the bithorax complex. The resulting animals fail to make the normal bxd noncoding RNA and show no transcription across the bxd Polycomb Response Element in early embryos. The mutant flies look normal; the regulation of the bxd domain appears unaffected. Thus, the bxd noncoding RNA has no apparent function.

Nanofibrillated cellulose composite hydrogel for the replacement of the nucleus pulposus.

The swelling and compressive mechanical behavior as well as the morphology and biocompatibility of composite hydrogels based on Tween® 20 trimethacrylate (T3), N-vinyl-2-pyrrolidone (NVP) and nanofibrillated cellulose (NFC) were assessed in the present study. The chemical structure of T3 was verified by Fourier transform infrared spectroscopy and proton nuclear magnetic resonance, and the degree of substitution was found to be around 3. Swelling ratios of neat hydrogels composed of different concentrations of T3 and NVP were found to range from 1.5 to 5.7 with decreasing concentration of T3. Various concentrations of cellulose nanofibrils (0.2-1.6wt.%) were then used to produce composite hydrogels that showed lower swelling ratios than neat ones for a given T3 concentration. Neat and composite hydrogels exhibited a typical nonlinear response under compression. All composite hydrogels showed an increase in elastic modulus compared to neat hydrogel of about 3- to 8-fold, reaching 18kPa at 0% strain and 62kPa at 20% strain for the hydrogel with the highest NFC content. All hydrogels presented a porous and homogeneous structure, with interconnected pore cells of around 100nm in diameter. The hydrogels are biocompatible. The results of this study demonstrate that composite hydrogels reinforced with NFC may be viable as nucleus pulposus implants due to their adequate swelling ratio, which may restore the annulus fibrosus loading, and their increased mechanical properties, which could possibly restore the height of the intervertebral discs.

Goosecoid and cerberus-like do not interact during mouse embryogenesis.

Mouse Cerberus-like (Cer-l) is a neural inducer molecule, capable of inhibiting Nodal and BMP-4 signals in the extracelular space. The cer-l expression domain in the Anterior Visceral Endoderm (AVE) and prechordal plate, tissues involved in head induction and patterning, respectively, suggested a role for this gene in head formation. However, animals homozygous for the cer-l null allele failed to show any abnormality, leading us to propose the existence of other factor(s) that might compensate for cer-l loss-of-function. Since goosecoid (gsc) shares some domains of expression with cer-l and was shown to be essential for head morphogenesis, we tested its ability to interact genetically with cer-l. With this aim we generated cer-l;gsc double mutants. These animals were analyzed at birth for skeletal defects and revealed the same phenotype as gsc-/- single mutants. We also investigated the proper patterning of structures adjacent to the prechordal plate by performing in situ hybridization of HNF-3beta, Six-3 and BF-1, genes whose expression domains remained unchanged. In conclusion, the analysis carried out indicated that gsc does not compensate for cer-l loss-of-function and that these genes do not interact genetically.