RANKL-induced DC-STAMP is essential for osteoclastogenesis

Osteoclasts are bone-resorbing, multinucleated giant cells that are essential for bone remodeling and are formed through cell fusion of mononuclear precursor cells. Although receptor activator of nuclear factor– B ligand (RANKL) has been demonstrated to be an important osteoclastogenic cytokine, the cell surface molecules involved in osteoclastogenesis are mostly unknown. Here, we report that the seven-transmembrane receptor-like molecule, dendritic cell–specific transmembrane protein (DC-STAMP) is involved in osteoclastogenesis. Expression of DCSTAMP is rapidly induced in osteoclast precursor cells by RANKL and other osteoclastogenic stimulations. Targeted inhibition of DC-STAMP by small interfering RNAs and specific antibody markedly suppressed the formation of multinucleated osteoclast-like cells. Overexpression of DC-STAMP enhanced osteoclastogenesis in the presence of RANKL. Furthermore, DC-STAMP directly induced the expression of the osteoclast marker tartrate-resistant acid phosphatase. These data demonstrate for the first time that DC-STAMP has an essential role in osteoclastogenesis.Toshio Kukita, Naohisa Wada, Akiko Kukita, Takashi Kakimoto, Ferry Sandra, Kazuko Toh, Kengo Nagata, Tadahiko Iijima, Madoka Horiuchi, Hiromi Matsusaki, Kunio Hieshima, Osamu Yoshie and Hisayuki Nomiyam

Constraints on Triton atmospheric evolution from occultations: 1989-2022

Context - Around the year 2000, Triton's south pole experienced an extreme summer solstice that occurs every about 650 years, when the subsolar latitude reached about 50{\deg}. Bracketing this epoch, a few occultations probed Triton's atmosphere in 1989, 1995, 1997, 2008 and 2017. A recent ground-based stellar occultation observed on 6 October 2022 provides a new measurement of Triton's atmospheric pressure which is presented here. Aims- The goal is to constrain the Volatile Transport Models (VTMs) of Triton's atmosphere that is basically in vapor pressure equilibrium with the nitrogen ice at its surface. Methods - Fits to the occultation light curves yield Triton's atmospheric pressure at the reference radius 1400 km, from which the surface pressure is induced. Results - The fits provide a pressure p_1400= 1.211 +/- 0.039 microbar at radius 1400 km (47 km altitude), from which a surface pressure of p_surf= 14.54 +/- 0.47 microbar is induced (1-sigma error bars). To within error bars, this is identical to the pressure derived from the previous occultation of 5 October 2017, p_1400 = 1.18 +/- 0.03 microbar and p_surf= 14.1 +/- 0.4 microbar, respectively. Based on recent models of Triton's volatile cycles, the overall evolution over the last 30 years of the surface pressure is consistent with N2 condensation taking place in the northern hemisphere. However, models typically predict a steady decrease in surface pressure for the period 2005-2060, which is not confirmed by this observation. Complex surface-atmosphere interactions, such as ice albedo runaway and formation of local N2 frosts in the equatorial regions of Triton could explain the relatively constant pressure between 2017 and 2022.Comment: 8 pages, 4 figures, accepted for publication in Astronomy and Astrophysic

Mouse anti-RANKL antibody delays oral wound healing and increases TRAP-positive mononuclear cells in bone marrow

Objectives: Denosumab, a human monoclonal antibody (mAb) that neutralizes receptor activator for nuclear factor κB ligand (RANKL), is associated with osteonecrosis of the jaw. However, the effect of denosumab on oral wounds is unclear. The aim was to determine the effect of anti-RANKL mAb on oral wounds and bone marrow. Materials and methods: The direct effect of the mAb on fibroblasts, macrophages, and osteoclasts were assessed in vitro. In vivo, mouse anti-RANKL mAb was administered to mice for 9 weeks prior to palatal bone denudation surgery. Mice were euthanized 3 weeks post-surgery, and wound healing was histomorphometrically analyzed. Long bones were assessed using micro-computed tomography, quantitative real-time polymerase chain reaction, and flow cytometry. Results: The mAb had no effect on macrophages and fibroblasts but significantly suppressed osteoclast proliferation in vitro. The mAb treatment significantly increased bone mass by suppressing osteoclasts in vivo. The expression of pro-osteoclastic genes was promoted in the bone marrow of the mAb-administered animals. Consistently, the mAb significantly induced the development of tartrate-resistant acid phosphatase (TRAP)-positive mononuclear cells (MNCs) but not osteoclasts in bone marrow. The mAb treatment had no effect on gross healing of the palatal wounds. However, significant inflammation was retained in the connective tissue facing the once denuded bone surface. Conclusions: Repair of the damaged palate was delayed, and significant inflammation was sustained in the connective tissue by anti-RANKL mAb treatment. Clinical relevance: Denosumab impairs osteoclastic bone repair. Care should be exercised to minimize osseous trauma when invasive procedures are performed on patients taking denosumab

A computational method for estimating the PCR duplication rate in DNA and RNA-seq experiments

BACKGROUND: PCR amplification is an important step in the preparation of DNA sequencing libraries prior to high-throughput sequencing. PCR amplification introduces redundant reads in the sequence data and estimating the PCR duplication rate is important to assess the frequency of such reads. Existing computational methods do not distinguish PCR duplicates from “natural” read duplicates that represent independent DNA fragments and therefore, over-estimate the PCR duplication rate for DNA-seq and RNA-seq experiments. RESULTS: In this paper, we present a computational method to estimate the average PCR duplication rate of high-throughput sequence datasets that accounts for natural read duplicates by leveraging heterozygous variants in an individual genome. Analysis of simulated data and exome sequence data from the 1000 Genomes project demonstrated that our method can accurately estimate the PCR duplication rate on paired-end as well as single-end read datasets which contain a high proportion of natural read duplicates. Further, analysis of exome datasets prepared using the Nextera library preparation method indicated that 45–50% of read duplicates correspond to natural read duplicates likely due to fragmentation bias. Finally, analysis of RNA-seq datasets from individuals in the 1000 Genomes project demonstrated that 70–95% of read duplicates observed in such datasets correspond to natural duplicates sampled from genes with high expression and identified outlier samples with a 2-fold greater PCR duplication rate than other samples. CONCLUSIONS: The method described here is a useful tool for estimating the PCR duplication rate of high-throughput sequence datasets and for assessing the fraction of read duplicates that correspond to natural read duplicates. An implementation of the method is available at https://github.com/vibansal/PCRduplicates. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12859-017-1471-9) contains supplementary material, which is available to authorized users