Particulate
matter (PM) air pollution poses a formidable public
health threat to the city of Beijing. Among the various hazards of
PM pollutants, microorganisms in PM2.5 and PM10 are thought to be responsible for various allergies and for the
spread of respiratory diseases. While the physical and chemical properties
of PM pollutants have been extensively studied, much less is known
about the inhalable microorganisms. Most existing data on airborne
microbial communities using 16S or 18S rRNA gene sequencing to categorize
bacteria or fungi into the family or genus levels do not provide information
on their allergenic and pathogenic potentials. Here we employed metagenomic
methods to analyze the microbial composition of Beijing’s PM
pollutants during a severe January smog event. We show that with sufficient
sequencing depth, airborne microbes including bacteria, archaea, fungi,
and dsDNA viruses can be identified at the species level. Our results
suggested that the majority of the inhalable microorganisms were soil-associated
and nonpathogenic to human. Nevertheless, the sequences of several
respiratory microbial allergens and pathogens were identified and
their relative abundance appeared to have increased with increased
concentrations of PM pollution. Our findings may serve as an important
reference for environmental scientists, health workers, and city planners.
Spermatogenesis is a differentiation process during which diploid spermatogonial stem cells (SSCs) produce haploid spermatozoa. This highly specialized process is precisely controlled at the transcriptional, posttranscriptional, and translational levels. Here we report that N-methyladenosine (mA), an epitranscriptomic mark regulating gene expression, plays essential roles during spermatogenesis. We present comprehensive mA mRNA methylomes of mouse spermatogenic cells from five developmental stages: undifferentiated spermatogonia, type A spermatogonia, preleptotene spermatocytes, pachytene/diplotene spermatocytes, and round spermatids. Germ cell-specific inactivation of the mA RNA methyltransferase Mettl3 or Mettl14 with Vasa-Cre causes loss of mA and depletion of SSCs. mA depletion dysregulates translation of transcripts that are required for SSC proliferation/differentiation. Combined deletion of Mettl3 and Mettl14 in advanced germ cells with Stra8-GFPCre disrupts spermiogenesis, whereas mice with single deletion of either Mettl3 or Mettl14 in advanced germ cells show normal spermatogenesis. The spermatids from double-mutant mice exhibit impaired translation of haploid-specific genes that are essential for spermiogenesis. This study highlights crucial roles of mRNA mA modification in germline development, potentially ensuring coordinated translation at different stages of spermatogenesis.
Worldwide, myopia is the leading cause of visual impairment. It results from inappropriate extension of the ocular axis and concomitant declines in scleral strength and thickness caused by extracellular matrix (ECM) remodeling. However, the identities of the initiators and signaling pathways that induce scleral ECM remodeling in myopia are unknown. Here, we used single-cell RNA-sequencing to identify pathways activated in the sclera during myopia development. We found that the hypoxia-signaling, the eIF2-signaling, and mTOR-signaling pathways were activated in murine myopic sclera. Consistent with the role of hypoxic pathways in mouse model of myopia, nearly one third of human myopia risk genes from the genome-wide association study and linkage analyses interact with genes in the hypoxia-inducible factor-1α (HIF-1α)-signaling pathway. Furthermore, experimental myopia selectively induced HIF-1α up-regulation in the myopic sclera of both mice and guinea pigs. Additionally, hypoxia exposure (5% O) promoted myofibroblast transdifferentiation with down-regulation of type I collagen in human scleral fibroblasts. Importantly, the antihypoxia drugs salidroside and formononetin down-regulated HIF-1α expression as well as the phosphorylation levels of eIF2α and mTOR, slowing experimental myopia progression without affecting normal ocular growth in guinea pigs. Furthermore, eIF2α phosphorylation inhibition suppressed experimental myopia, whereas mTOR phosphorylation induced myopia in normal mice. Collectively, these findings defined an essential role of hypoxia in scleral ECM remodeling and myopia development, suggesting a therapeutic approach to control myopia by ameliorating hypoxia.
Near-ultraviolet nitride-based light-emitting diodes (LEDs) with peak emission wavelengths around 410 nm were fabricated onto c-face patterned sapphire substrates (PSS). It was found that the electroluminescence intensity of the PSS LED shown 63% larger than that of the conventional LED. For a typical lamp-form PSS LED operating at a forward current of 20 mA, the output power and external quantum efficiency were estimated to be 10.4 mW and 14.1%, respectively. The improvement in the light intensity could be attributed to the decrease of threading dislocations and the increase of light extraction efficiency in the horizontal direction using a PSS
Metagenomic sequencing has been widely used for the study of microbial communities from various environments such as soil, ocean, sediment and fresh water. Nonetheless, metagenomic sequencing of microbial communities in the air remains technically challenging, partly owing to the limited mass of collectable atmospheric particulate matter and the low biological content it contains. Here we present an optimized protocol for extracting up to tens of nanograms of airborne microbial genomic DNA from collected particulate matter. With an improved sequencing library preparation protocol, this quantity is sufficient for downstream applications, such as metagenomic sequencing for sampling various genes from the airborne microbial community. The described protocol takes ∼12 h of bench time over 2-3 d, and it can be performed with standard molecular biology equipment in the laboratory. A modified version of this protocol may also be used for genomic DNA extraction from other environmental samples of limited mass or low biological content.
Even though there were slight percentage changes between contact lens wearers and nonwearers in some microbes, there were no differences in their diversity. On the other hand, contact lens usage might cause relative abundance of some taxa to change. Our results will help assess whether or not conjunctival microbiome changes caused by contact lens wear affect infection risk.
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