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Zygotic genome activation (ZGA) is a critical developmental milestone characterized by the rapid and simultaneous activation of genome-widely silenced chromatin. Various active histone modifications accumulate upon ZGA and have long been implicated in ZGA. However, their biological relevance still remains unclear. Here, we comprehensively examined the functional significance of active histone modifications and their writers during ZGA in teleost embryos. Our data propose that developmental genes and housekeeping genes are distinctively regulated during ZGA; CBP/P300 activity is required for developmental gene activation, whereas housekeeping genes depend on non-CBP/P300 histone acetylations H3K9ac/H4K16ac/H3K14ac. Moreover, accumulation of H3K4me2/3 is not prerequisite for activation of all types of genes during ZGA, in contrast to previous reports with cell lines. Finally, temporal accumulation of H3.3S31ph greatly enhances CBP/P300 activity specifically at ZGA, ensuring the activation of developmental genes. Our data demonstrate that multiple histone modifications cooperatively shape ZGA-specific gene activation programs in non-mammalian vertebrates. The role of active histone modifications during zygotic genome activation (ZGA) is not fully understood in vertebrates. Here, the authors propose two types of activation mediated by different histone modifications during ZGA in teleost embryos.

Inter/transgenerational epigenetic inheritance is a crucial and controversial theory that could reshape the concept of genetics. To investigate this theory directly, we invent a system for targeted reprogramming of epigenetic memory in mouse sperm. Using this system, we erase DNA methylation at the differentially methylated region of the H19 gene ( H19 -DMR) in sperm, which causes Silver-Russell syndrome-like phenotypes in F1 offspring. Although DNA methylation is fully lost in the sperm, it is partially restored during pre-implantation development, suggesting the existence of epigenetic memory that instructs de novo DNA methylation. Importantly, targeted removal of histone modifications in zygotes reveals that tri-methylation at lysine 9 of histone H3 (H3K9me3), which is deposited shortly after fertilization, is required for the subsequent de novo DNA methylation at the H19 -DMR. Thus, our study provides a robust germline editing tool, which reveals partial intergenerational inheritance and no transgenerational inheritance at the model locus. Furthermore, we identify H3K9me3 as a mediator for DNA methylation recovery also acting at imprinted loci. The authors develop a system to erase DNA methylation at H19 -DMR in mouse sperm. Despite loss, methylation partly recovers via H3K9me3 after fertilization, revealing partial intergenerational epigenetic inheritance.

This report describes generation of dendritic cells (DCs) and macrophages from human induced pluripotent stem (iPS) cells. iPS cell-derived DC (iPS-DC) exhibited the morphology of typical DC and function of T-cell stimulation and antigen presentation. iPS-DC loaded with cytomegalovirus (CMV) peptide induced vigorous expansion of CMV-specific autologous CD8 + T cells. Macrophages (iPS-MP) with activity of zymosan phagocytosis and C5a-induced chemotaxis were also generated from iPS cells. Genetically modified iPS-MPs were generated by the introduction of expression vectors into undifferentiated iPS cells, isolation of transfectant iPS cell clone and subsequent differentiation. By this procedure, we generated iPS-MP expressing a membrane-bound form of single chain antibody (scFv) specific to amyloid β (Aβ), the causal protein of Alzheimer’s disease. The scFv-transfectant iPS-MP exhibited efficient Aβ-specific phagocytosis activity. iPS-MP expressing CD20-specific scFv engulfed and killed BALL-1 B-cell leukemia cells. Anti-BALL-1 effect of iPS-MP in vivo was demonstrated in a xeno-transplantation model using severe combined immunodeficient mice. In addition, we established a xeno-free culture protocol to generate iPS-DC and iPS-MP. Collectively, we demonstrated the possibility of application of iPS-DC and macrophages to cell therapy.

Forages serve an important role in providing nutrients to ruminants while providing positive benefits to the environment. Forage cell wall digestibility is incomplete because of several structural features within the wall, but digestion is mostly inversely correlated with the amount of lignification that has occurred during cell wall development. Lignin is a hydrophobic polymer formed through enzyme-mediated radical coupling of monolignols, mainly coniferyl and sinapyl alcohols. The polymer is highly resistant to degradation and generally passes through the ruminant unmodified. Though lignin is resistant to degradation, it is not easily quantified within various types of forages. Numerous methods have been developed over the years to measure lignin levels in different plant species. Most frequently used among workers involved with forage development or utilization are the acid detergent, Klason, and permanganate lignin methods. More recently, acetyl bromide has received attention as a possible lignin determination method. The acetyl bromide method is dependent on determining the absorbance of the extract in which all the lignin of a sample has been dissolved. Each of these methods gives different lignin values for the same type of forage sample. For example, acid detergent, Klason, permanganate, and acetyl bromide lignin methods give quite different values for alfalfa stems: 93, 145, 158, and 135 g lignin kg(-1) cell wall, respectively. These differences can be even greater for grasses: 25, 77, 45, and 92 g kg(-1) cell wall from corn (Zea mays L.) stalks analyzed by acid detergent, Klason, permanganate, and acetyl bromide lignin methods, respectively. This paper will discuss the different lignin determination methods and highlight the advantages and disadvantages of each as they relate to forage sample analysis.

Background Epigenetic modifications have a central role in transcriptional regulation. While several studies using next-generation sequencing have revealed genome-wide associations between epigenetic modifications and transcriptional states, a direct causal relationship at specific genomic loci has not been fully demonstrated, due to a lack of technology for targeted manipulation of epigenetic modifications. Recently, epigenome editing techniques based on the CRISPR-Cas9 system have been reported to directly manipulate specific modifications at precise genomic regions. However, the number of editable modifications as well as studies applying these techniques in vivo is still limited. Results Here, we report direct modification of the epigenome in medaka (Japanese killifish, Oryzias latipes ) embryos. Specifically, we developed a method to ectopically induce the repressive histone modification, H3K27me3 in a locus-specific manner, using a fusion construct of Oryzias latipes H3K27 methyltransferase Ezh2 (olEzh2) and dCas9 (dCas9-olEzh2). Co-injection of dCas9-olEzh2 mRNA with single guide RNAs (sgRNAs) into one-cell-stage embryos induced specific H3K27me3 accumulation at the targeted loci and induced downregulation of gene expression. Conclusion In this study, we established the in vivo epigenome editing of H3K27me3 using medaka embryos. The locus-specific manipulation of the epigenome in living organisms will lead to a previously inaccessible understanding of the role of epigenetic modifications in development and disease.

Wildlife trade is a major driver of biodiversity loss, yet whilst the impacts of trade in some species are relatively well-known, some taxa, such as many invertebrates are often overlooked. Here we explore global patterns of trade in the arachnids, and detected 1,264 species from 66 families and 371 genera in trade. Trade in these groups exceeds millions of individuals, with 67% coming directly from the wild, and up to 99% of individuals in some genera. For popular taxa, such as tarantulas up to 50% are in trade, including 25% of species described since 2000. CITES only covers 30 (2%) of the species potentially traded. We mapped the percentage and number of species native to each country in trade. To enable sustainable trade, better data on species distributions and better conservation status assessments are needed. The disparity between trade data sources highlights the need to expand monitoring if impacts on wild populations are to be accurately gauged and the impacts of trade minimised. Trade in arachnids includes millions of individuals and over 1264 species, with over 70% of individuals coming from the wild.

Background: Myxoid liposarcoma (MLS) is the second most common subtype of liposarcoma, and metastasis occurs in up to one-third of cases. However, the mechanisms of invasion and metastasis remain unclear. Tumour-associated macrophages (TAMs) have important roles in tumour invasion, metastasis, and/or poor prognosis. The aim of this study was to investigate the relationship between TAMs and MLS. Methods: Using 78 primary MLS samples, the association between clinical prognosis and macrophage infiltration was evaluated by immunochemistry. The effects of macrophages on cell growth, cell motility, and invasion of MLS cell lines were investigated in vitro . In addition, clinicopathological factors were analysed to assess their prognostic implications in MLS. Results: Higher levels of CD68-positive macrophages were associated with poorer overall survival in MLS samples. Macrophage-conditioned medium enhanced MLS cell motility and invasion by activating epidermal growth factor receptor (EGFR), with the key ligand suggested to be heparin-binding EGF-like growth factor (HB-EGF). The phosphoinositide 3-kinase/Akt pathway was mostly involved in HB-EGF-induced cell motility and invasion of MLS. The expression of phosphorylated EGFR in MLS clinical samples was associated with macrophage infiltration. In addition, more significant macrophage infiltration was associated with poor prognosis even in multivariate analysis. Conclusions: Macrophage infiltration in MLS predicts poor prognosis, and the relationship between TAMs and MLS may be a new candidate for therapeutic targets of MLS.

Background:The rate of elderly people over 65 year-old increased from 22.1 % in 2008 to 27.7% in 2017 in Japan, also from 27.1 % to 32.3 % in our super-aging area1, 2. The number of total and unilateral knee arthroplasty (TKA, UKA) have increased annually in all over the world according to the larger population of elderly people due to osteoarthritis (OA)3. In fact, the numbers of primary TKA predicted increasing from six hundred fifty-six thousand cases at 2010 to one million three hundred seventy-six thousand cases at 2020 in USA4. In the other hand, rheumatoid arthritis (RA) therapy have been remarkably improved from starting to use biologic agents since 2003 in Japan 5. The rate of orthopaedic surgery may reflect trends in disease severity and drug management of RA 5.Objectives:The aim of study is to reveal the rate of TKA, including UKA and revision TKA in elderly people in our super-aging area of Japan.Methods:We surveyed the number and cause of primary and revision TKA and UKA in our institutes using the data of diagnosis procedure combination and the record of surgeries in the last decade.Results:Figure 1.Table 1.2008-122013-17TimesOsteoarthritis15652252*1.3Rheumatoid arthritis13181*0.6Trauma155Osteonecrosis of femoral condylar2041*2.1Revision21391.9Total17382418*1.4* p< 0.05Conclusion:The number and rate of primary TKA/UKA due to RA decreased year by year because of progression of modern medication therapy. In the other hand, in case of OA increased because of increasing of elderly people affected by knee OA in the super-aging society.References:[1]National Institute of Population and Social Security Research. Japanese Mortality Database, 2018. http://www.ipss.go.jp/[2]Yamagata prefecture, Health and longevity Promotion Section. Rate of elderly people in Yamagata prefecture, 2018. http://www.pref.yamagata.jp/ou/kikakushinko/020052/tokei/jinkel.html[3]Annual report 2017 of replacement arthroplasty in Japan. The Japanese Society For Replacement Arthroplasty The Japan Arthroplasty Register. https://jsra.info/jar-report.html[4]Kurtz SM, et al. J Bone Joint Surg Am. 96: 624-30, 2014.[5]Momohara S, et al. J Rheumatol. 41:862-5, 2014.Disclosure of Interests:None declared

Background The extinction of species is a multifaceted phenomenon shaped by the complex interplay between biological and socio-cultural factors. Public and academic preferences for different species often play a direct or indirect role in influencing the conservation outlook of these species. The “charisma” of species and other components of biodiversity is often mentioned as an important factor in shaping human preferences, determining both the scope of scientific studies and justifications for such scope. Here, we present a protocol for systematically mapping the use of the concept of “charisma” in relation to biodiversity peer-reviewed academic literature focused on biodiversity conservation. Methods The search targeting academic peer-reviewed research articles and reviews will be conducted in three publication databases, The Lens, Scopus and Web of Science (Core Collection and SciELO), and will be supplemented by search engine results from Google Scholar. Broad-scope searches will be performed in 3 different languages (English, Portuguese, and Spanish) and article screening will be performed at two stages to ensure the relevance of each entry and consistency amongst reviewers in their use of the defined inclusion criteria. The resulting systematic map of the literature will be summarised by employing a narrative synthesis approach, and through descriptive statistics and analysis of temporal trends.

This paper presents a new correlative bioimaging technique using Y 2 O 3 :Tm, Yb and Y 2 O 3 :Er, Yb nanophosphors (NPs) as imaging probes that emit luminescence excited by both near-infrared (NIR) light and an electron beam. Under 980 nm NIR light irradiation, the Y 2 O 3 :Tm, Yb and Y 2 O 3 :Er, Yb NPs emitted NIR luminescence (NIRL) around 810 nm and 1530 nm, respectively and cathodoluminescence at 455 nm and 660 nm under excitation of accelerated electrons, respectively. Multimodalities of the NPs were confirmed in correlative NIRL/CL imaging and their locations were visualized at the same observation area in both NIRL and CL images. Using CL microscopy, the NPs were visualized at the single-particle level and with multicolour. Multiscale NIRL/CL bioimaging was demonstrated through in vivo and in vitro NIRL deep-tissue observations, cellular NIRL imaging and high-spatial resolution CL imaging of the NPs inside cells. The location of a cell sheet transplanted onto the back muscle fascia of a hairy rat was visualized through NIRL imaging of the Y 2 O 3 :Er, Yb NPs. Accurate positions of cells through the thickness (1.5 mm) of a tissue phantom were detected by NIRL from the Y 2 O 3 :Tm, Yb NPs. Further, locations of the two types of NPs inside cells were observed using CL microscopy.