The Fab portion of immunoglobulin G contributes to its binding to Fcγ receptor III

Most cells active in the immune system express receptors for antibodies which mediate a variety of defensive mechanisms. These receptors interact with the Fc portion of the antibody and are therefore collectively called Fc receptors. Here, using high-speed atomic force microscopy, we observe interactions of human, humanized, and mouse/human-chimeric immunoglobulin G1 (IgG1) antibodies and their cognate Fc receptor, FcγRIIIa. Our results demonstrate that not only Fc but also Fab positively contributes to the interaction with the receptor. Furthermore, hydrogen/deuterium exchange mass spectrometric analysis reveals that the Fab portion of IgG1 is directly involved in its interaction with FcγRIIIa, in addition to the canonical Fc-mediated interaction. By targeting the previously unidentified receptor-interaction sites in IgG-Fab, our findings could inspire therapeutic antibody engineering

Targeted reversion of induced pluripotent stem cells from patients with human cleidocranial dysplasia improves bone regeneration in a rat calvarial bone defect model

BackgroundRunt-related transcription factor 2 (RUNX2) haploinsufficiency causes cleidocranial dysplasia (CCD) which is characterized by supernumerary teeth, short stature, clavicular dysplasia, and osteoporosis. At present, as a therapeutic strategy for osteoporosis, mesenchymal stem cell (MSC) transplantation therapy is performed in addition to drug therapy. However, MSC-based therapy for osteoporosis in CCD patients is difficult due to a reduction in the ability of MSCs to differentiate into osteoblasts resulting from impaired RUNX2 function. Here, we investigated whether induced pluripotent stem cells (iPSCs) properly differentiate into osteoblasts after repairing the RUNX2 mutation in iPSCs derived from CCD patients to establish normal iPSCs, and whether engraftment of osteoblasts derived from properly reverted iPSCs results in better regeneration in immunodeficient rat calvarial bone defect models.MethodsTwo cases of CCD patient-derived induced pluripotent stem cells (CCD-iPSCs) were generated using retroviral vectors (OCT3/4, SOX2, KLF4, and c-MYC) or a Sendai virus SeVdp vector (KOSM302L). Reverted iPSCs were established using programmable nucleases, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas-derived RNA-guided endonucleases, to correct mutations in CCD-iPSCs. The mRNA expressions of osteoblast-specific markers were analyzed using quantitative reverse-transcriptase polymerase chain reaction. iPSCs-derived osteoblasts were transplanted into rat calvarial bone defects, and bone regeneration was evaluated using microcomputed tomography analysis and histological analysis.ResultsMutation analysis showed that both contained nonsense mutations: one at the very beginning of exon 1 and the other at the initial position of the nuclear matrix-targeting signal. The osteoblasts derived from CCD-iPSCs (CCD-OBs) expressed low levels of several osteoblast differentiation markers, and transplantation of these osteoblasts into calvarial bone defects created in rats with severe combined immunodeficiency showed poor regeneration. However, reverted iPSCs improved the abnormal osteoblast differentiation which resulted in much better engraftment into the rat calvarial bone defect.ConclusionsTaken together, these results demonstrate that patient-specific iPSC technology can not only provide a useful disease model to elucidate the role of RUNX2 in osteoblastic differentiation but also raises the tantalizing prospect that reverted iPSCs might provide a practical medical treatment for CCD

Diatom biostratigraphy in sediments of ODP Sites 204-1251 and 204-1252

Sites 1251 (44°34.213'N, 125°4.440'W; 1211 m water depth) and 1252 (44°35.167'N, 125°5.569'W; 1039 m water depth) were drilled on the eastern flank of the southern summit of Hydrate Ridge off Oregon in the northeast Pacific Ocean, where well-stratified sediments were deposited at a rapid rate. Unconformities and debris flow layers of middle Pleistocene age were found at both sites. Their ages are of great importance in constructing the geohistory of Hydrate Ridge. Detailed diatom biostratigraphy of the middle to late Pleistocene of Sites 1251 and 1252 was carried out for this purpose

Crown beaked whale fossils from the Chepotsunai Formation (latest Miocene) of Tomamae Town, Hokkaido, Japan

In the last decades, our knowledge of ziphiid evolution has increased dramatically. However, their periotic morphology is still poorly known. A fossil ziphiid (TTM-1) including the periotic, bulla, isolated polydont teeth and vertebrae from the Chepotsunai Formation (latest Miocene) of Tomamae Town, Hokkaido, Japan, is identified as a member of a clade with crown ziphiids of Bianucci et al. (2016) by having three periotic synapomorphies; a posteriorly wide posterior process, transversely thick anterior process, and laterally elongated lateral process. The specimen adds morphological information of the periotic. Among the Ziphiidae from the stem to crown, the periotic morphologies were changed to having a more robust anterior process, wider anterior bullar facet and posterior process. The crown Ziphiidae shares a feature; enlarged medial tubercle on the anterior process. Among the crown Ziphiidae, TTM-1 does not have a swollen medial tubercle not like Tasmacetus, Nazcacetus and others. This new morphological information might represent useful future phylogenetic comparisons