Terashima, A. | bioRxiv Science

bioRxiv2022

intra-single cell sequencing (iSCseq) spotlights transcriptomic and epigenetic heterogeneity inside multinucleated osteoclast

Single-cell RNA-seq (scRNA-seq) has clarified cellular heterogeneity within cell populations. However, scRNA-seq and spatial transcriptomics cannot capture the dynamic transcriptomic changes inside living cells. To decode subcellular gene expression, we developed intra-single cell sequencing (iSCseq), a novel approach that combines confocal imaging, repeatedly picking up cellular components inside living cells, and next-generation sequencing (intra single-cell RNA-seq; iSCseq). iSCseq illustrated the subcellular heterogeneity of gene expression. iSCseq revealed not only multiple differentiation stages embedded in the same cell, but also physical cytoskeletal connections, physiological activity of mitochondria, and intracellular calcium, as confirmed by transcriptomic evidence. Inclusive iSCseq with in vivo scRNA-seq datasets identified new osteoclast subsets in physiological and pathological bones. Network analysis with centrality provided insights into the connection between subcellular components, and clearly divided differentiation and fusion processes in multinucleation. The iSCseq approach has the potential to enhance cell biology at subcellular resolution and identify new therapeutic targets. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/506360v4_ufig1.gif" ALT="Figure 1"> View larger version (73K): org.highwire.dtl.DTLVardef@115c1d8org.highwire.dtl.DTLVardef@1695ad2org.highwire.dtl.DTLVardef@1c9d257org.highwire.dtl.DTLVardef@1107ddb_HPS_FORMAT_FIGEXP M_FIG C_FIG In briefintra-single cell sequencing (iSCseq) enhances single-cell technology by combining live cell imaging, subcellular sampling from living cells and sequencing, offering deeper insights into cell functions and pathology at subcellular resolution through inclusive analysis with scRNA-seq and advanced centrality-focused network analysis. HighlightsO_LIintra-single cell sequencing (iSCseq) clarifies subcellular heterogeneity C_LIO_LIiSCseq connects morphological and physiological features with transcriptome C_LIO_LIInclusive iSCseq unveils osteoclast subsets in physiological and pathological bones C_LIO_LILinkage at subcellular resolution reveals key players in characteristic fusion C_LI

genomics↗

bioRxiv2018

Soil-transmitted helminthiasis in children of a rural community part of a school-based deworming program: a cross-sectional study in the Peruvian Amazon

BackgroundChildren in the Peruvian Amazon basin are at risk of soil-transmitted helminthiases (STH). The aim of this study was to determine the prevalence of STH (Ascaris lumbricoides, Trichuris trichiura, hookworm and Strongyloides stercoralis) in children from a rural community in the Peruvian Amazon and associated clinical, maternal, sanitation and hygiene factors associated. The community had an active school-based deworming program with mebendazole.\n\nMethodsA cross-sectional study was carried out in children aged 2-14 years in Iquitos, Peru; by parasitological analysis of their stools through five methods: direct smear examination, Kato-Katz, spontaneous sedimentation in tube, Baermann method modified by Lumbreras and agar plate culture. Mothers of the participating children were also invited to participate in the study. A questionnaire was completed by each participant to collect demographic and epidemiological information of interest.\n\nResultsAmong 124 children, 25.8% (32/124) had one or more STH. Prevalence of A. lumbricoides was 16.1% (20/124); S. stercoralis, 10.5% (13/124); hookworm, 1.6% (2/124) and T. trichiura, 1.6% (2/124). STH in mothers was higher in children with any STH than in children without any STH (36.4% vs 14.1%, p<0.02). Prevalence of the common STH (A. lumbricoides, T. trichiura and hookworm) was higher in children aged 2-5 than in older children (31.6% vs 12.8%; p=0.01). Several hygiene and sanitation deficits were identified; of which walking barefoot was significantly associated with STH infection (OR= 3.28; 95% CI= 1.11-12.07).\n\nConclusionsSTH are highly prevalent in children in this community; A. lumbricoides and S. stercoralis infections were the most frequent. Further studies should aim to understand the persistent high prevalence of common STH in endemic areas where massive drug administration is practiced, and to determine the appropriateness of controlling STH in mothers and S. stercoralis infection. Walking barefoot and other hygiene and sanitation conditions need to be addressed in this community.\n\nAUTHOR SUMMARYFew studies assessing soil-transmitted helminth infections in children and their risk factors have yet been conducted in the Peruvian Amazon. Even fewer reports exist from areas where mass drug administration programs have been initiated. In this study we provide insight to the frequency of soil-transmitted helminths in a setting with an ongoing school-based deworming program.\n\nBesides the most common soil-transmitted helminths (Ascaris lumbricoides, Trichuris trichiura and hookworm), we assessed the prevalence of Strongyloides stercoralis. Excluding the latter from intestinal helminths studies have often underestimate its frequency and impact in children.\n\nWe also surveyed for helminth infection in the mothers of the participating children as infection in caregivers could theoretically be associated with infection in children as they both may share same environmental and behavioral factors associated with STH infections. To our knowledge, this is the first Peruvian study assessing children and mother infection together.\n\nIn addition, our results highlight the suboptimal hygiene and sanitation conditions in which people from this rural community live. It likely represents the situation of other rural Amazonian communities in Peru. Public efforts are needed to provide these populations with dignified living conditions and to follow the equity global health agenda.

epidemiology↗