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Current pharmacological treatments for endometriosis are limited to hormonal agents that can relieve pain but cannot cure the disease. Therefore, the development of a disease-modifying drug for endometriosis is an unmet medical need. By studying human endometriotic samples, we found that the progression of endometriosis was associated with the development of inflammation and fibrosis. In addition, IL-8 expression was highly up-regulated in endometriotic tissues and closely correlated with disease progression. We created a long-acting recycling antibody against IL-8 (AMY109) and evaluated its clinical potency. Because rodents do not produce IL-8 and do not experience menstruation, we analyzed the lesions in cynomolgus monkeys that spontaneously developed endometriosis and in a surgically induced endometriosis monkey model. Both spontaneously developed and surgically induced endometriotic lesions demonstrated pathophysiology that was highly similar to that of human endometriosis. Once-a-month subcutaneous injection of AMY109 to monkeys with surgically induced endometriosis reduced the volume of nodular lesions, lowered the Revised American Society for Reproductive Medicine score as modified for monkeys, and ameliorated fibrosis and adhesions. In addition, experiments using cells derived from human endometriosis revealed that AMY109 inhibited the recruitment of neutrophils to endometriotic lesions and the production of monocyte chemoattractant protein-1 from neutrophils. Thus, AMY109 may represent a disease-modifying therapy for patients with endometriosis.

Dysregulation of the complement system is linked to the pathogenesis of a variety of hematological disorders. Eculizumab, an anti-complement C5 monoclonal antibody, is the current standard of care for paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic uremic syndrome (aHUS). However, because of high levels of C5 in plasma, eculizumab has to be administered biweekly by intravenous infusion. By applying recycling technology through pH-dependent binding to C5, we generated a novel humanized antibody against C5, SKY59, which has long-lasting neutralization of C5. In cynomolgus monkeys, SKY59 suppressed C5 function and complement activity for a significantly longer duration compared to a conventional antibody. Furthermore, epitope mapping by X-ray crystal structure analysis showed that a histidine cluster located on C5 is crucial for the pH-dependent interaction with SKY59. This indicates that the recycling effect of SKY59 is driven by a novel mechanism of interaction with its antigen and is distinct from other known pH-dependent antibodies. Finally, SKY59 showed neutralizing effect on C5 variant p.Arg885His, while eculizumab does not inhibit complement activity in patients carrying this mutation. Collectively, these results suggest that SKY59 is a promising new anti-C5 agent for patients with PNH and other complement-mediated disorders.

Various studies have demonstrated that Fc engineering to enhance neonatal Fc receptor (FcRn) binding is effective for elongating half-life or increasing cellular uptake of IgG. A previous study has shown that a N434H mutation to enhance FcRn binding resulted in increased binding to rheumatoid factor (RF) autoantibody, which is not desirable for therapeutic use in autoimmune disease. In this study, we first showed that all the existing Fc variants with enhanced FcRn binding also show increased RF binding, and then identified specific mutations that could be introduced to those Fc variants to reduce the RF binding. Furthermore, we generated novel Fc variants that do not increase RF binding and show half-lives of 45 d in cynomolgus monkey, which is longer than those of previously reported Fc variants. In addition, we generated novel Fc variants with antigen sweeping activity that do not increase RF binding. We expect that these novel Fc variants will be useful as antibody therapeutics against autoimmune diseases.

Previously, we have reported novel engineered antibody with pH-dependent antigen-binding (recycling antibody), and with both pH-dependent antigen-binding and increased FcRn-binding at neutral pH (sweeping antibody). The purpose of this study is to perform PK/PD predictions to better understand the potential applications of the antibodies as therapeutics. To demonstrate the applicability of recycling and sweeping antibodies over conventional antibodies, PK/PD analyses were performed. PK/PD parameters for antibody and antigen dynamics were estimated from the results of a pharmacokinetic study in human FcRn transgenic mice. A simulation study was performed using the estimated PK/PD parameters with various target antigen profiles. In comparison to conventional antibody, recycling antibody enhanced antibody-antigen complex clearance by 3 folds, while sweeping antibody accelerated antigen clearance by 10 folds in a pharmacokinetic study. Simulation results showed that recycling and sweeping antibodies can improve dosage frequency and reduce the required dose for target antigens with various clearances, plasma concentrations or binding kinetics. Moreover, importance of the association rate constant to enhance the beneficial effect of antibodies was shown. These results support the conclusion that recycling and sweeping antibodies can be applied to various target antigens with different profiles, and expand the number of antigens that antibodies can target.

Immunogenicity is a key factor capable of influencing the efficacy and safety of therapeutic antibodies. A recently developed method called MHC-associated peptide proteomics (MAPPs) uses liquid chromatography/mass spectrometry to identify the peptide sequences derived from a therapeutic protein that are presented by major histocompatibility complex class II (MHC II) on antigen-presenting cells, and therefore may induce immunogenicity. In this study, we developed a MAPPs technique (called Ab-MAPPs) that has high throughput and can efficiently identify the MHC II-presented peptides derived from therapeutic antibodies using magnetic nanoparticle beads coated with a hydrophilic polymer in the immunoprecipitation process. The magnetic beads could identify more peptides and sequence regions originating from infliximab and adalimumab in a shorter measurement time than Sepharose beads, which are commonly used for MAPPs. Several sequence regions identified by Ab-MAPPs from infliximab corresponded to immunogenic sequences reported by other methods, which suggests the method’s high potential for identifying significant sequences involved in immunogenicity. Furthermore, our study suggests that the Ab-MAPPs method can recognize the difference of a single amino acid residue between similar antibody sequences with different levels of T-cell proliferation activity and can identify potentially immunogenic peptides with high binding affinity to MHC II. In conclusion, Ab-MAPPs is useful for identifying the immunogenic sequences of therapeutic antibodies and will contribute to the design of therapeutic antibodies with low immunogenicity during the drug discovery stage.

Heart rate monitoring has huge potential in disease prevention, stroke prediction, and mental stress/workload assessment. Although most conventional heart rate monitoring systems are wearable devic...

We demonstrate a newly-developed process for replicating a surface shape of an atomically smooth master substrate onto electroplated Au patterns by a lift-off process using a thin sacrificial layer. In this process, a Ti sacrificial layer and a thin Au seed layer were first deposited on the master substrate. Sealing ring patterns were then formed using a combination of photolithography and Au electroplating. These patterns were next bonded to a Au thin film on a Si wafer. Finally, by chemically dissolving the Ti sacrificial layer, the patterns were released from the master substrate and transferred to the Si wafer. The resulting patterns had an atomically smooth surface with a root-mean-square surface roughness of 0.8 nm. To examine applicability of the smooth Au surface to Au–Au direct bonding, these smooth patterns were bonded to another Au-coated Si wafer at room temperature in atmospheric air, and tensile tests were carried out. A high bonding strength of about 250 MPa was obtained, with fracture eventually occurring within the Si substrate.

Rough surfaces of electroplated Au patterns were flattened by thermal imprinting.Almost the entire area of a 100-µm-wide pattern was flattened.Large bonding strength was obtained by using a flattening process.Narrow multi-wall line patterns were designed for the bonding strength and hermeticity.Bonding strength was further improved by flattening and a multi-wall narrow pattern. This paper reports on the flattening of a rough surface of an electroplated Au pattern on a Si wafer by the thermal imprint process and its application to surface-activated room-temperature bonding. An ultra-flat sapphire wafer was pressed onto the rough surface of an electroplated Au pattern on a Si wafer for 10min at 200?C with an applied pressure of 150MPa. As a result, almost entire area of the 100-µm-wide pattern was flattened. Flattened Au patterns on Si wafer were bonded to a Au thin film on a Si wafer by surface-activated room-temperature bonding. A large bonding strength above 200MPa was obtained. Narrow multi-wall line patterns were designed in order to ensure the bonding strength and hermeticity. The bonding strength was further improved by a combination of flattening and the multi-wall narrow pattern.

Graphical abstractDisplay Omitted We evaluated the characterization of bonding interface between Si surfaces smoothed by Ne FAB surface treatment.The rough surface with a surface roughness of 0.40nm rms was finally decreased down to 0.17nm rms by Ne FAB etching.With Ne FAB smoothing, a bonding energy equivalent to that of the bulk materials was finally attained.Local strain at the bonded interface between surfaces was dramatically decreased by Ne FAB smoothing. We performed room-temperature Si wafer direct bonding by surface activation using a Ne fast atom beam (FAB). The bonding energy between Si wafers prepared by Ne FAB etching is equivalent to that of the bulk materials. We also investigated the surface smoothing effect by Ne FAB etching. A rough Si surface with a surface roughness of 0.40nm rms was etched by Ne FAB, and its surface roughness was decreased down to 0.17nm rms. For a Si surface etched at a depth of 30nm by the Ne FAB, the 1D-PSD (1-dimensional power spectrum density) amplitude was decreased in the range of spatial frequencies >10µm-1. Although the Si wafers with surface roughness of 0.4nm rms could not be bonded, they were successfully bonded after Ne FAB smoothing of etching depths over 5nm, and a bonding energy equivalent to that of the bulk materials was attained. We succeeded in dramatically decreasing the local strain at the bonding interface by the Ne FAB smoothing effect.

We developed a new room temperature wafer bonding of metal films using flattening by thermal imprint process.Au film surfaces were successfully flattened with a 5MPa pressure at 200?C for 600s.Developed room temperature bonding was effective when the bonding load is small and/or the Au films are relatively rough. This paper reports on the surface flattening of metal films on wafers by the thermal imprint process and its application to wafer direct bonding. The ultraflat surface of a sapphire wafer was pressed onto the surface of a sputter-deposited Au film on a Si wafer for 600s at 200?C with an applied pressure of 5MPa by compressed air. As a result, the Au film surface was successfully flattened by thermal imprint process. The flattened Au films were bonded in a vacuum chamber after surface activation by Ar beam sputtering. In the case of bonding between two Au films on a Si wafer, the surface flattening of the Au films was determined to be effective when the bonding load is small and/or the Au films are relatively thick. In the case of bonding between a Si wafer and a Au film on a Si wafer, defects at the bonding interface were found to be dramatically reduced by the flattening of the Au film surface. Additionally, the Au-Si bonding using the flattened Au film was significantly stronger than that without flattening.

Japan is aging rapidly, so "tojikomori" has become an important issue. In the field of gerontology, "tojikomori" means shutting oneself in a room or house. Tojikomori is caused not only by physical factors but also by social and psychological ones. Once an elderly person becomes tojikomori, it takes a lot of time and energy to recover. It is important to take primary preventive measures, especially for currently active older adults because a little help has a great benefit. A previous study identified two key factors of tojikomori in an urban area: no exercise habits and no participation in community activities. Therefore, we developed a walking game to enhance physically healthy behavior and motivate the elderly to explore a city and lead to the discovery of favorite spots they want to visit again. In addition, we designed a group-walking program utilizing the walking game aimed for face-to-face communication with other community-dwelling elderly people. Our results show that our approach was well received, enjoyable, and effective for older adults in a real-world situation.

We evaluated room temperature bonding characteristics of electroplated Au surfaces smoothed by lift-off method and imprint method. As a result, smoothed surfaces can make a strong bonding; on the other hand electroplated rough surface make a very weak bonding. In TEM observation, no delamination was observed at the bonding interface, which was bonded at room temperature using a smooth surface replicated by the lift-off process. Hermeticity of bonding interface prepared by smoothed surfaces was evaluated by diaphragm structures. As a result, good hermetic sealing was confirmed using the smooth surface replicated by the lift-off method.

We demonstrate a newly-developed replication process of a surface shape from an atomically smooth master substrate onto electroplated Au patterns by a lift-off process using a thin sacrificial layer. An atomically smooth Au surface with a root mean square surface roughness of 0.8 nm could be replicated from the master substrate by this process. Then we examined its applicability to room-temperature Au-Au bonding in atmosphere. A high bonding strength of about 250 MPa was obtained. Fracture from the bulk of the Si substrates was observed after tensile tests, which was also the case for the thermocompression bonding at 200 °C.

Capturing the user's vital signs is an urgent goal in the HCI community. Photo-plethysmography (PPG) is one approach; it can collect data from the finger tips that indicate the user's autonomic nervous system (ANS) and offers new potentials such as mental stress measurement and drowsy state detection. Our goal is to set PPG sensors on the surfaces of ordinary devices such as mice, smartphones, and steering wheels. This will offer smart monitoring without the burden of additional wearable sensors. Unfortunately, current PPG sensors are very narrow, and even if the sensor is attached to the surface of a device, the user is forced to align and hold the finger to the sensor point, which degrades device usability. To solve this problem, we propose an area-based sensing method that relaxes the alignment requirement. The proposed method uses two thin acrylic plates, a diffuser plate and a detection plate, as an IR waveguide. The proposed method can yield very thin sensing surfaces and gentle curvatures are possible. An experiment compares the proposed method to the conventional point-sensor in terms of LF/HF discrimination performance with the participant in the resting state, and the proposed method is shown to offer comparable sensing performance with superior usability.

For many antibodies, each antigen-binding site binds to only one antigen molecule during the antibody's lifetime in plasma. To increase the number of cycles of antigen binding and lysosomal degradation, we engineered tocilizumab (Actemra), an antibody against the IL-6 receptor (IL-6R), to rapidly dissociate from IL-6R within the acidic environment of the endosome (pH 6.0) while maintaining its binding affinity to IL-6R in plasma (pH 7.4). Studies using normal mice and mice expressing human IL-6R suggested that this pH-dependent IL-6R dissociation within the acidic environment of the endosome resulted in lysosomal degradation of the previously bound IL-6R while releasing the free antibody back to the plasma to bind another IL-6R molecule. In cynomolgus monkeys, an antibody with pH-dependent antigen binding, but not an affinity-matured variant, significantly improved the pharmacokinetics and duration of C-reactive protein inhibition. Engineering pH dependency into the interactions of therapeutic antibodies with their targets may enable them to be delivered less frequently or at lower doses.

Fc engineering to increase the binding affinity of IgG antibodies to FcRn has been reported to reduce the elimination of IgG antibodies. Herein, we present a novel non-FcRn-dependent approach to reduce the elimination of IgG antibodies. Pharmacokinetic studies conducted in normal mice of various humanized IgG4 antibodies, which had identical constant regions but different variable region sequences, revealed that an antibody with a lower isoelectric point (pI) has a longer half-life. These antibodies exhibited comparable binding affinity to FcRn, and with the antibodies with lower pIs, a longer half-life was also observed in beta2-microglobulin knockout mice, suggesting that differences in the pharmacokinetics were due to a non-FcRn-dependent mechanism. On the basis of our findings, we attempted to engineer the pharmacokinetic properties of a humanized anti-IL6 receptor IgG1 antibody. Selected substitutions in the variable region, without substitution in the Fc region, lowered the pI but did not reduce the biological activity and showed a significant reduction in the clearance of the antibody in cynomolgus monkey. These results suggest that lowering the pI by engineering the variable region could reduce the elimination of IgG antibodies and could provide an alternative to Fc engineering of IgG antibodies.

We evaluated room-temperature bonding characteristics of electroplated Au surfaces smoothed by the lift-off and imprint methods. As a result, we found that smoothed surfaces enable strong bonding; on the other hand, electroplated rough surfaces result in very weak bonding. In transmission electron microscopy observations, no delamination was observed at the bonding interface bonded at room temperature using a smooth surface prepared by the lift-off method. Moreover, the hermeticity of the bonding interface prepared using smoothed surfaces was evaluated using diaphragm structures. As a result, we confirmed that good hermetic sealing was achieved using the electroplated Au surface smoothed by the lift-off method.

We have developed a wafer-scale layer-transfer technique for transferring GaAs and Ge onto Si wafers of up to 300 mm in diameter. Lattice-matched GaAs or Ge layers were epitaxially grown on GaAs wafers using an AlAs release layer, which can subsequently be transferred onto a Si handle wafer via direct wafer bonding and patterned epitaxial lift-off (ELO). The crystal properties of the transferred GaAs layers were characterized by X-ray diffraction (XRD), photoluminescence, and the quality of the transferred Ge layers was characterized using Raman spectroscopy. We find that, after bonding and the wet ELO processes, the quality of the transferred GaAs and Ge layers remained the same compared to that of the as-grown epitaxial layers. Furthermore, we realized Ge-on-insulator and GaAs-on-insulator wafers by wafer-scale pattern ELO technique.

This poster describes an implementation of the puzzle generators and puzzle layouts introduced in [1].

Ge and III-V compound semiconductors are expected to be used for non-Si channel MOSFETs as one of the effective technical booster to reduce the power consumption of next generation VLSI because of its higher carrier mobility than that of Si. Various approaches to introduce those non-Si channel materials into Si platform have been proposed, among which the ELO(epitaxial lift off) technique may have a good potential for manufacturing such a composite wafer consist of non-lattice-matched crystalline materials. High quality Ge and GaAs/InGaAs epitaxial thin film can be grown on lattice-matched/quasi-lattice-matched GaAs or InP single crystal substrate using CVD/MOCVD technology. The thin epitaxial film can be chemically lifted-off by selectively removing sacrificial epitaxial layer grown between the epitaxial film and the substrate, transferred and directly bonded onto Si substrate using thin oxide film made of Al2O3 and/or SiO2 as an adhesive layer, thus successively providing GOI and/or III-VOI(Ge and/or III-V compound semiconductors on insulator) composite wafer including the oxide film as an effective BOX(buried oxide) layer. The area size of transferred Ge/III-V film can be modified according to its application die size. The epitaxial layer quality after transferred onto Si substrate and initial device performance of Ge/III-V channel MOSFET formed on the Ge/III-V OI substrate will be introduced.

In the surface activated bonding (SAB) at room temperature, the surfaces of specimens are sputter etched by high-energy ion/atom beam of inert gas to remove surface contaminants and create active surfaces, which are ready to form inter-atomic bonds even at room temperature. Ar has been used for the purpose. However, sputter etching by Ar high-energy ion/atom beam causes increase of surface roughness. In case of wafer direct bonding, the surface roughness has large influence on the quality of bonding interface such as bonding strength, local strain, and so on. Therefore a surface activation process which does not cause surface roughening has been desired. In this work, we have examined the applicability of Ne fast-atom-beam (FAB) to SAB of Si wafers at room temperature. We confirmed that Ne-FAB does not cause surface roughening and Si wafers are successfully bonded by Ne-beam surface activation. In addition, we found that Ne-FAB sputter etching has surface smoothing effect which improves quality of the bonding interface. The room temperature wafer bonding apparatus (MWB-12-ST: Mitsubishi Heavy Industries) with the FAB 104 (Atom Tech Ltd.) beam sources were used for bonding of Si wafers. The etching rate of Si surface by Ne-FAB was about half of that of Ar-FAB using the beam source operation parameters of 1.8 kV applied voltage and 100 mA input current. With etching depth more than 1.5 nm, the bonding strength comparable to Si bulk strength was attained by Ne-FAB surface activation. With etching depth of 30 nm, we measured surface roughening of Si wafers using Ne, Ar and Xe gas. The surface roughness of Si wafers largely increased by the Ar and Xe-FAB sputter etching. In case of Ne-FAB, the surface roughness did not increase by the sputter etching and kept the initial rms value of 0.18 nm of CMP-polished Si wafers. Moreover, we found surface smoothing effect of the Ne-FAB. We prepared Si wafers which had been roughened by Xe-FAB etching. They could not be bonded directly by SAB. Surface roughness of the roughened Si wafer decreased from 0.40 to 0.17 nm rms by applying Ne-FAB of 30 nm etching depth. Along with the surface smoothing, the bonding strength was largely improved and finally became equivalent to Si bulk strength. It was confirmed that strain near the bonding interface was decreased by the smoothing effect as shown in Figure.

We examined the applicability of a Ne fast atom beam (FAB) to surface activated bonding of Si wafers at room temperature. With etching depth more than 1.5 nm, the bonding strength comparable to Si bulk strength was attained. Moreover, we found the improvement of the bonding strength by surface smoothing effect of the Ne FAB. Silicon surface roughness decreased from 0.40 to 0.17 nm rms by applying a Ne FAB of 30 nm etching depth. The bonding strength between surfaces recovered by Ne FAB surface smoothing was largely improved and finally became equivalent to Si bulk strength.

Monoclonal antibodies are widely used to target disease-related antigens. However, because conventional antibody binds to the antigen but cannot eliminate the antigen from plasma, and rather increases the plasma antigen concentration by reducing the clearance of the antigen, some clinically important antigens are still difficult to target with monoclonal antibodies because of the huge dosages required. While conventional antibody can only bind to the antigen, some natural endocytic receptors not only bind to the ligands but also continuously eliminate them from plasma by pH-dependent dissociation of the ligands within the acidic endosome and subsequent receptor recycling to the cell surface. Here, we demonstrate that an engineered antibody, named sweeping antibody, having both pH-dependent antigen binding (to mimic the receptor-ligand interaction) and increased binding to cell surface neonatal Fc receptor (FcRn) at neutral pH (to mimic the cell-bound form of the receptor), selectively eliminated the antigen from plasma. With this novel antigen-sweeping activity, antibody without in vitro neutralizing activity exerted in vivo efficacy by directly eliminating the antigen from plasma. Moreover, conversion of conventional antibody with in vitro neutralizing activity into sweeping antibody further potentiated the in vivo efficacy. Depending on the binding affinity to FcRn at neutral pH, sweeping antibody reduced antigen concentration 50- to 1000-fold compared to conventional antibody. Thereby, sweeping antibody antagonized excess amounts of antigen in plasma against which conventional antibody was completely ineffective, and could afford marked reduction of dosage to a level that conventional antibody can never achieve. Thus, the novel mode of action of sweeping antibody provides potential advantages over conventional antibody and may allow access to the target antigens which were previously undruggable by conventional antibody.

Thermal conduction in a one-dimensional anharmonic lattice is investigated with the use of homogeneous nonequilibrium molecular dynamics. This approach enables us to obtain thermal conductivity as a function of temperature. Coherent excitations are observed to play an important role in transporting energy, especially when an external ``field'' becomes large.

We have identified a partial deletion of the short arm of chromosome 20 in a 5-year-old boy from parents having normal phenotype and karyotype. His major anomalies were mild mental retardation, congenital heart disease, chest deformity, spina bifida, kyphoscoliosis, inguinal hernia, and preauricular fistula. The clinical findings were compared with those of two patients reported previously as a partial deletion 20p.

We have realized patterned Ge-on-Insulator wafers by large-scale layer transfer technology. In conjunction with low-temperature bonding and patterned Epitaxial Lift-Off (ELO) technique, high quality Ge layer transfer was achieved in full-wafer scale.