Review: Insects—A Source of Safe and Sustainable Food?— “Jein” (Yes and No)

For almost a decade, edible insects have become promoted on a wider basis as one way to combat world hunger and malnourishment, although attempts to do so have a longer history. Contemporary researchers and consumers, particularly those without an entomophagous background, have been rising safety and sustainability concerns. The present contribution seeks a substantiated answer to the question posed above. The possible answer consists of different factors that have been taken into consideration. First, the species and its life cycle. It is mandatory to realize that what is labeled as “edible insects” stands for more than 2,140 animal species, not counting other edible, non-crustacean arthropods. Their life cycles are as diverse as the ecological niches these animals can fill and last between some days to several years and many of them may—or may not—be reproduced in the different farming systems. Second, the level of knowledge concerning the food use of a given species is important, be it traditional, newly created by research, or a combination of both. Third, the existence of a traditional method of making the use of the insect safe and sustainable, ideally from both the traditional and the modern points of view. Fourth, the degree of effectiveness of these measures despite globalization changes in the food-supplying network. Fifth, farming conditions, particularly housing, feeding (type, composition, and contaminants), animal health and animal welfare. Sixth, processing, transport, and storage conditions of both traditional and novel insect-based foodstuffs, and seventh, consumer awareness and acceptance of these products. These main variables create a complex web of possibilities, just as with other foodstuffs that are either harvested from the wild or farmed. In this way, food safety may be reached when proper hygiene protocols are observed (which usually include heating steps) and the animals do not contain chemical residues or environment contaminants. A varying degree of sustainability can be achieved if the aforementioned variables are heeded. Hence, the question if insects can be safe and sustainable can be answered with “jein,” a German portmanteau word joining “yes” (“ja”) and “no” (“nein”).

Divergent evolution of developmental timing in the neocortex revealed by marsupial and eutherian transcriptomes

Only mammals evolved a neocortex, which integrates sensory-motor and cognitive functions. Significant diversifications in the cellular composition and connectivity of the neocortex occurred between the two main Therian groups: marsupials and eutherians. However, the developmental mechanisms underlying these diversifications are largely unknown. Here, we compared the neocortical transcriptomes of Sminthopsis crassicaudata, a mouse-sized marsupial, with those of eutherian mice at two developmentally equivalent timepoints corresponding to deeper and upper layer neuron generation. Enrichment analyses revealed more mature gene networks in marsupials at the early stage, which reverted at the later stage, suggesting a more precocious but protracted neuronal maturation program relative to birth timing of cortical layers. We ranked genes expressed in different species and identified important differences in gene expression rankings between species. For example, genes known to be enriched in upper-layer cortical projection neuron subtypes, such as Cux1, Lhx2 and Satb2, likely relating to corpus callosum emergence in eutherians. These results show molecular heterochronies of neocortical development in Theria, and highlight changes in gene expression and cell type composition that may underlie neocortical evolution and diversification.

Postnatal developmental trajectory of sex-biased gene expression in the mouse pituitary gland

The pituitary gland controls sexually dimorphic processes such as growth, pubertal onset, and lactation. However, the mechanisms underlying sex biases in pituitary gene regulation are not fully understood. To capture pituitary gene regulation dynamics during postnatal development, we ascertained gene and miRNA expression across five postnatal days that span the pubertal transition in mice. Using three prime untranslated region and small RNA sequencing, we observed over 900 instances of sex-biased gene expression, including 18 genes that were putative targets of 5 sex-biased miRNAs. In addition, by combining bulk RNA-seq with scRNA-seq pituitary data, we obtained evidence that cell-type proportion sex differences exist prior to puberty and contribute substantially to the observed sex-biased gene expression post-puberty. This work provides a resource for postnatal mouse pituitary gene regulation and highlights the importance of sex-biases in both cell-type composition and gene regulation when understanding the sexually dimorphic processes regulated by the pituitary gland.

DNA Methylation Profiles of Purified Cell Types in Bronchoalveolar Lavage: Applications for Mixed Cell Paediatric Pulmonary Studies

In epigenome-wide association studies analysing DNA methylation from samples containing multiple cell types, it is essential to adjust the analysis for cell type composition. One well established strategy for achieving this is reference-based cell type deconvolution, which relies on knowledge of the DNA methylation profiles of purified constituent cell types. These are then used to estimate the cell type proportions of each sample, which can then be incorporated to adjust the association analysis. Bronchoalveolar lavage is commonly used to sample the lung in clinical practice and contains a mixture of different cell types that can vary in proportion across samples, affecting the overall methylation profile. A current barrier to the use of bronchoalveolar lavage in DNA methylation-based research is the lack of reference DNA methylation profiles for each of the constituent cell types, thus making reference-based cell composition estimation difficult. Herein, we use bronchoalveolar lavage samples collected from children with cystic fibrosis to define DNA methylation profiles for the four most common and clinically relevant cell types: alveolar macrophages, granulocytes, lymphocytes and alveolar epithelial cells. We then demonstrate the use of these methylation profiles in conjunction with an established reference-based methylation deconvolution method to estimate the cell type composition of two different tissue types; a publicly available dataset derived from artificial blood-based cell mixtures and further bronchoalveolar lavage samples. The reference DNA methylation profiles developed in this work can be used for future reference-based cell type composition estimation of bronchoalveolar lavage. This will facilitate the use of this tissue in studies examining the role of DNA methylation in lung health and disease.

Single-cell transcriptomic landscapes of a rare human laryngeal chondrosarcoma

Propose Laryngeal chondrosarcoma is a rare non-epithelial malignant tumor. At present, the cell type composition and molecular mechanism of laryngeal chondrosarcoma have not been systematically studied. Methods This study focused on the histopathological and imaging features of a rare primary laryngeal chondrosarcoma in a 74-year-old male. The tumor and its paracancerous cartilage tissue were single-cell sequenced and analyzed and a total of 5455 single cells were obtained. Immunohistochemical levels were also verified. Results In total five cell types were identified, including chondrocytes, myeloid cells, fibroblasts, lymphocytes, and endothelial cells. We carried out further subgroup analysis, focusing on the classification and differentiation of chondrocytes, functional enrichment analysis, and cellular communication analysis of all cell types, and explored the tumor microenvironment (TME) of laryngeal chondrosarcoma. Immunohistochemistry revealed the SLAMF9 gene was specifically expressed in non-immune cells of chondrosarcoma, but was barely expressed in the normal cartilage tissues adjacent to chondrosarcomas. Conclusion This single-cell sequencing approach provides clues for deciphering the potential mechanisms of tumor heterogeneity and TME composition in laryngeal chondrosarcoma, and represents an important step towards the treatment of laryngeal chondrosarcoma.

Lower promoter activity of the ST8SIA2 gene has been favored in evolving human collective brains

ST8SIA2 is an important molecule regulating expression of the phenotype involved in schizophrenia. Lowered promoter activity of the ST8SIA2 gene is considered to be protective against schizophrenia by conferring tolerance to psychosocial stress. Here, we examined the promoter-type composition of anatomically modern humans (AMHs) and archaic humans (AHs; Neanderthals and Denisovans), and compared the promoter activity at the population level (population promoter activity; PPA) between them. In AMHs, the TCT-type, showing the second lowest promoter activity, was most prevalent in the ancestral population of non-Africans. However, the detection of only the CGT-type from AH samples and recombination tracts in AH sequences showed that the CGT- and TGT-types, exhibiting the two highest promoter activities, were common in AH populations. Furthermore, interspecies gene flow occurred into AMHs from AHs and into Denisovans from Neanderthals, influencing promoter-type compositions independently in both AMHs and AHs. The difference of promoter-type composition makes PPA unique in each population. East and Southeast Asian populations show the lowest PPA. This results from the selective increase of the CGC-type, showing the lowest promoter activity, in these populations. Every non-African population shows significantly lower PPA than African populations, resulting from the TCT-type having the highest prevalence in the ancestral population of non-Africans. In addition, PPA reduction is also found among subpopulations within Africa via a slight increase of the TCT-type. These findings indicate a trend toward lower PPA in the spread of AMHs, interpreted as a continuous adaptation to psychosocial stress arising in migration. This trend is considered as genetic tuning for the evolution of collective brains. The inferred promoter-type composition of AHs differed markedly from that of AMHs, resulting in higher PPA in AHs than in AMHs. This suggests that the trend toward lower PPA is a unique feature in AMH spread.

Regenerated crustacean limbs are precise replicas

Animals can regenerate complex organs, yet this frequently results in imprecise replicas of the original structure. In the crustacean Parhyale, embryonic and regenerating legs differ in gene expression dynamics but produce apparently similar mature structures. We examine the fidelity of Parhyale leg regeneration using complementary approaches to investigate microanatomy, sensory function, cellular composition and cell molecular profiles. We find that regeneration precisely replicates the complex microanatomy and spatial distribution of external sensory organs, and restores their sensory function. Single-nuclei sequencing shows that regenerated and uninjured legs are indistinguishable in terms of cell type composition and transcriptional profiles. This remarkable fidelity highlights the ability of organisms to achieve identical outcomes via distinct processes.

Systematic evaluation of cell-type deconvolution pipelines for sequencing-based bulk DNA methylomes

DNA methylation sequencing is becoming increasingly popular, yielding genome-wide methylome data at single-base pair resolution through the novel cost- and labor-optimized protocols. It has tremendous potential for cell-type heterogeneity analysis, particularly in tumors, due to intrinsic read-level information. Although diverse deconvolution methods were developed to infer cell-type composition based on bulk sequencing-based methylomes, their systematic evaluation has not been performed so far. Here, we thoroughly review and evaluate five previously published deconvolution methods: Bayesian epiallele detection (BED), PRISM, csmFinder + coMethy, ClubCpG and MethylPurify, together with two array-based methods, MeDeCom and Houseman as a comparison group. Sequencing-based deconvolution methods consist of two main steps, informative region selection and cell-type composition estimation. Accordingly, we individually assessed the performance of each step and demonstrated the impact of the former step upon the performance of the following one. In conclusion, we demonstrate the best method showing the highest accuracy in different samples, and infer factors affecting cell-type deconvolution performance according to the number of cell types in the mixture. We found that cell-type deconvolution performance is influenced by different factors according to the number of components in the mixture. Whereas selecting similar genomic regions to DMRs generally contributed to increasing the performance in bi-component mixtures, the uniformity of cell-type distribution showed a high correlation with the performance in five cell-type bulk analyses.