Developmental competence of interspecies cloned embryos produced using cells from large Japanese field mice (Apodemus speciosus) and oocytes from laboratory mice (Mus musculus domesticus).

The large Japanese field mouse (Apodemus speciosus) is endemic to Japan and may be used as an animal model for studies related to environmental pollution, medical science, and basic biology. However, the large Japanese field mouse has low reproductive ability due to the small number of oocytes ovulated per female. To produce experimental models, we investigated the in vitro developmental potential of interspecies somatic cell nuclear transfer (iSCNT) embryos produced by fusing tail tip cells from the large Japanese field mouse with enucleated oocytes from laboratory mice (Mus musculus domesticus). Only a small number of iSCNT embryos developed to the 4-cell (0-4%) and blastocysts (0-1%) stages under sequential treatment using trichostatin A (TSA) and vitamin C (VC) supplemented with deionized bovine serum albumin (d-BSA). This sequential treatment led to the reduction in H3K9 trimethylation and did not affect H3K4 trimethylation in at least the 2-cell stage of the iSCNT embryos. Moreover, iSCNT embryos that received tail tip cells with exposure treatment to ooplasm from cell fusion to oocyte activation or VC treatment prior to cell fusion did not exhibit significant in vitro development improvement compared to that of each control group. This suggests that large Japanese field mice/laboratory mice iSCNT embryos that received sequential treatment using TSA and VC with d-BSA may have slightly better developmental potential beyond the 4-cell stage. Our results provide insights into the reprogramming barriers impeding the wider implementation of iSCNT technology.

WDM photon pair source stabilized in a wide temperature range with cascaded optical nonlinearities and DFG monitoring.

In this paper, we report a multi-channel wavelength division multiplexed (WDM) photon pair source operating in a wide temperature range. The photon pair generation rates in multiple WDM channels were stabilized against changes in the operation temperature by combining the spectral flatness of cascaded optical nonlinearities (cascaded sum frequency generation/spontaneous parametric downconversion) with a differential frequency generation monitoring feedback system. The proposed method was experimentally validated using a type-I periodically poled LiNbO3 ridge waveguide device as the photon pair source. We successfully generated 16 WDM photon pairs at almost the same rate (0.024358 ± 0.000631 pairs/s/Hz), even when the operating temperature was varied from 27.01 °C to 60.16 °C.

Generation of highly stable WDM time-bin entanglement by cascaded sum-frequency generation and spontaneous parametric downconversion in a PPLN waveguide device.

In this paper we report the generation of wavelength-division-multiplexed, time-bin entangled photon pairs by using cascaded optical second nonlinearities (sum-frequency generation and subsequent spontaneous parametric downconversion) in a periodically poled LiNbO3 device. Visibilities of approximately 94% were clearly observed in two-photon interference experiments for all the wavelength-multiplexed channels under investigation (five pairs), with insensitivity to the polarization states of the photon pairs. We also evaluated the performances in terms of quantum-key-distribution (QKD) applications by using four single-photon detectors, which enables to evaluate the QKD performance properly. The results showed long-term stability over 70 hours, maintaining approximately 3% of the quantum error rate and 110 bit/s of the sifted key rate.

Highly efficient phase-sensitive parametric gain in periodically poled LiNbO3 ridge waveguide.

We have developed a periodically poled LiNbO3 (PPLN) ridge waveguide device and experimentally demonstrated a phase-sensitive optical parametric amplification. The highly efficient net phase-sensitive parametric gain of +18.6 dB is obtained in a cascaded second harmonic generation and difference frequency generation configuration. The phase-sensitive gain is greater by +5.8 dB in comparison with the phase-insensitive gain. We successfully confirmed the phase-sensitive amplification with a high gain in the PPLN ridge waveguide.

A unique hinge binder of extremely selective aminopyridine-based Mps1 (TTK) kinase inhibitors with cellular activity.

Mps1, also known as TTK, is a dual-specificity kinase that regulates the spindle assembly check point. Increased expression levels of Mps1 are observed in cancer cells, and the expression levels correlate well with tumor grade. Such evidence points to selective inhibition of Mps1 as an attractive strategy for cancer therapeutics. Starting from an aminopyridine-based lead 3a that binds to a flipped-peptide conformation at the hinge region in Mps1, elaboration of the aminopyridine scaffold at the 2- and 6-positions led to the discovery of 19c that exhibited no significant inhibition for 287 kinases as well as improved cellular Mps1 and antiproliferative activities in A549 lung carcinoma cells (cellular Mps1 IC50=5.3 nM, A549 IC50=26 nM). A clear correlation between cellular Mps1 and antiproliferative IC50 values indicated that the antiproliferative activity observed in A549 cells would be responsible for the cellular inhibition of Mps1. The X-ray structure of 19c in complex with Mps1 revealed that this compound retains the ability to bind to the peptide flip conformation. Finally, comparative analysis of the X-ray structures of 19c, a deamino analogue 33, and a known Mps1 inhibitor bound to Mps1 provided insights into the unique binding mode at the hinge region.

Wavelength conversion of incoherent light by sum-frequency generation.

In this paper, we reveal that some kinds of optical nonlinearities are further enhanced when incoherent light, instead of a laser, is used as a pump light. This idea was confirmed both theoretically and experimentally in the case of sum-frequency generation (SFG) using the optical second nonlinearity. The conversion efficiency of the SFG with incoherent light pumping increased as the bandwidth of the incoherent pump light decreased, finally reaching twice the conversion efficiency of conventional second harmonic generation (SHG) by laser pumping. This method dramatically relaxes the severe requirements of phase matching in the nonlinear optical process. The conversion efficiency became less sensitive to misalignment of the wavelength of pump light and also of device operation temperature when the bandwidth of the incoherent pump light was sufficiently broad, although the improvement of the conversion efficiency had an inverse relationship with the insensitivity to the phase-matching condition. The temperature tuning range was enhanced by more than two orders of magnitude in comparison with the conventional SHG method. As an example of a promising application of this new idea, we performed the generation of quantum entangled photon-pairs using cascaded optical nonlinearities (SFG and the subsequent spontaneous parametric down conversion) in a single periodically poled LiNbO3 waveguide device, in which the incoherent light was used as the pump source for both the parametric processes. We have achieved high fidelity exceeding 99% in quantum-state tomography experiments.

Carbamoyl pyridone HIV-1 integrase inhibitors 3. A diastereomeric approach to chiral nonracemic tricyclic ring systems and the discovery of dolutegravir (S/GSK1349572) and (S/GSK1265744).

We report herein the discovery of the human immunodeficiency virus type-1 (HIV-1) integrase inhibitors dolutegravir (S/GSK1349572) (3) and S/GSK1265744 (4). These drugs stem from a series of carbamoyl pyridone analogues designed using a two-metal chelation model of the integrase catalytic active site. Structure-activity studies evolved a tricyclic series of carbamoyl pyridines that demonstrated properties indicative of once-daily dosing and superior potency against resistant viral strains. An inherent hemiaminal ring fusion stereocenter within the tricyclic carbamoyl pyridone scaffold led to a critical substrate controlled diastereoselective synthetic strategy whereby chiral information from small readily available amino alcohols was employed to control relative and absolute stereochemistry of the final drug candidates. Modest to extremely high levels of stereochemical control were observed depending on ring size and position of the stereocenter. This approach resulted in the discovery of 3 and 4, which are currently in clinical development.

Indazole-based potent and cell-active Mps1 kinase inhibitors: rational design from pan-kinase inhibitor anthrapyrazolone (SP600125).

Monopolar spindle 1 (Mps1) is essential for centrosome duplication, the spindle assembly check point, and the maintenance of chromosomal instability. Mps1 is highly expressed in cancer cells, and its expression levels correlate with the histological grades of cancers. Thus, selective Mps1 inhibitors offer an attractive opportunity for the development of novel cancer therapies. To design novel Mps1 inhibitors, we utilized the pan-kinase inhibitor anthrapyrazolone (4, SP600125) and its crystal structure bound to JNK1. Our design efforts led to the identification of indazole-based lead 6 with an Mps1 IC50 value of 498 nM. Optimization of the 3- and 6-positions on the indazole core of 6 resulted in 23c with improved Mps1 activity (IC50 = 3.06 nM). Finally, application of structure-based design using the X-ray structure of 23d bound to Mps1 culminated in the discovery of 32a and 32b with improved potency for cellular Mps1 and A549 lung cancer cells. Moreover, 32a and 32b exhibited reasonable selectivities over 120 and 166 kinases, respectively.

Nearly degenerate wavelength-multiplexed polarization entanglement by cascaded optical nonlinearities in a PPLN ridge waveguide device.

In this paper we report the generation of wavelength-multiplexed polarization-entangled photon pairs in the 1.5-µm communication wavelength band by using cascaded optical second nonlinearities (sum-frequency generation and subsequent spontaneous parametric down-conversion, c-SFG/SPDC) in a periodically poled LiNbO(3) ridge waveguide device. The c-SFG/SPDC method makes it possible to fully use the broad spectral bandwidth of SPDC in nearly frequency-degenerate conditions, and can provide more than 50 pairs of wavelength channels for the entangled photon pairs in the 1.5-µm wavelength band, using only standard optical resources in the telecom field. Visibilities higher than 98% were clearly observed in two-photon interference fringes for all the wavelength channels under investigation (eight pairs). We further performed a detailed experimental investigation of the cross-talk characteristics and the impact of detuning the pump wavelengths.

Carbamoyl pyridone HIV-1 integrase inhibitors. 2. Bi- and tricyclic derivatives result in superior antiviral and pharmacokinetic profiles.

This work is a continuation of our initial discovery of a potent monocyclic carbamoyl pyridone human immunodeficiency virus type-1 (HIV-1) integrase inhibitor that displayed favorable antiviral and pharmacokinetic properties. We report herein a series of bicyclic carbamoyl pyridone analogues to address conformational issues from our initial SAR studies. This modification of the core unit succeeded to deliver low nanomolar potency in standard antiviral assays. An additional hydroxyl substituent on the bicyclic scaffold provides remarkable improvement of antiviral efficacies against clinically relevant resistant viruses. These findings led to additional cyclic tethering of the naked hydroxyl group resulting in tricyclic carbamoyl pyridone inhibitors to address remaining issues and deliver potential clinical candidates. The tricyclic carbamoyl pyridone derivatives described herein served as the immediate leads in molecules to the next generation integrase inhibitor dolutegravir which is currently in late stage clinical evaluation.

Carbamoyl pyridone HIV-1 integrase inhibitors. 1. Molecular design and establishment of an advanced two-metal binding pharmacophore.

Our group has focused on expanding the scope of a two-metal binding pharmacophore concept to explore HIV-1 integrase inhibitors through medicinal chemistry efforts to design novel scaffolds which allow for improvement of pharmacokinetic (PK) and resistance profiles. A novel chelating scaffold was rationally designed to effectively coordinate two magnesium cofactors and to extend an aromatic group into an optimal hydrophobic pharmacophore space. The new chemotype, consisting of a carbamoyl pyridone core unit, shows high inhibitory potency in both enzymatic and antiviral assay formats with low nM IC50 and encouraging potency shift effects in the presence of relevant serum proteins. The new inhibitor design displayed a remarkable PK profile suggestive of once daily dosing without the need for a PK booster as demonstrated by robust drug concentrations at 24 h after oral dosing in rats, dogs, and cynomolgus monkeys.

Experimental investigation in transmission performance of polarization-entangled photon-pairs generated by cascaded χ(2) processes over standard single-mode optical fibers.

In this paper we report experimental investigation in transmission performance over standard single-mode optical fibers (SMFs) of polarization-entangled photon-pairs in a 1.5-µm band generated by cascaded second-harmonic generation and spontaneous parametric down conversion (c-SHG/SPDC) from a periodically poled LiNbO(3) (PPLN) ridge-waveguide device. Clear two-photon interference fringes were observed even after the transmission over 140 km of the SMF spools, remaining small degradation in the visibilities of less than 3%. The performance was also investigated by using optical attenuators, instead of the SMF spools, to study the maximum reach of the distribution of the entanglement in terms of loss penalty. The results show that the quantum entanglement could be distributed even with 50 dB of the transmission loss with violation of Bell inequality by using the c-SHG/SPDC-based photon-pair source.

1.5-µm band polarization entangled photon-pair source with variable Bell states.

In this paper we report a polarization-entangled photon-pair source in a 1.5-µm band which can generate arbitrary entangled states including four maximum entangled states (Bell states) by using cascaded optical second nonlinearities (second-harmonic generation and the following spontaneous parametric down conversion) in a periodically poled LiNbO(3) (PPLN) ridge-waveguide device. Exchange among the Bell states was achieved by using an optical phase bias compensator (OPBC) in a Sagnac loop interferometer and a half-wave plate outside the loop for polarization conversion. Quantitative evaluation was made on the performance of the photon-pair source through the experiments of two-photon interferences, quantum state tomography, and test of violation of Bell inequality. We observed high visibilities of 96%, fidelities of 97%, and 2.71 of the S parameter in inequality of Clauser, Horne, Shimony, and Holt (CHSH). The experimental values, including peak coincidence counts in the two-photon interference (approximately 170 counts per second), remained almost unchanged in despite of the exchange among the Bell states. They were also in good agreement with the theoretical assumption from the mean number of the photon-pairs under the test (0.04 per pulse). More detailed experimental studies on the dependence of the mean number of the photon-pairs revealed that the quantum states were well understood as the Werner state.

Diaminopyridine-based potent and selective mps1 kinase inhibitors binding to an unusual flipped-Peptide conformation.

Monopolar spindle 1 (Mps1) is an attractive cancer drug target due to the important role that it plays in centrosome duplication, the spindle assembly checkpoint, and the maintenance of chromosomal stability. A design based on JNK inhibitors with an aminopyridine scaffold and subsequent modifications identified diaminopyridine 9 with an IC50 of 37 nM. The X-ray structure of 9 revealed that the Cys604 carbonyl group of the hinge region flips to form a hydrogen bond with the aniline NH group in 9. Further optimization of 9 led to 12 with improved cellular activity, suitable pharmacokinetic profiles, and good in vivo efficacy in the mouse A549 xenograft model. Moreover, 12 displayed excellent selectivity over 95 kinases, indicating the contribution of its unusual flipped-peptide conformation to its selectivity.

On-demand virtual optical network access using 100 Gb/s Ethernet.

Our Terabit LAN initiatives attempt to enhance the scalability and utilization of lambda resources. This paper describes bandwidth-on-demand virtualized 100GE access to WDM networks on a field fiber test-bed using multi-domain optical-path provisioning.

Monitoring technique for waveform distortion of 160 Gb/s signal by prescaled-clock tone detection using EA modulator.

In order to monitor quality of ultra high bit-rate optical signals in a future optical network, such as 160 Gb/s, a simple monitoring technique is required. Therefore, a novel waveform monitoring technique by prescaled-clock tone detection was proposed in a previous report. In this paper, detailed principle of the proposed technique was explained. The monitoring technique is based on an asynchronous beat signal generation using an elecro-absorption modulator (EAM) and is able to separately observe waveform distortion caused by accumulated chromatic dispersion (CD), polarization mode dispersion (PMD) and optical signal-to-noise ratio (OSNR) degradation. The verification of concepts was performed by experiments, in which 1 GHz pre-scaled signals were employed to monitor distortion of OTDM 160 Gb/s carrier suppressed return-to-zero (CS-RZ) signals. Furthermore, applicability to Q factor estimation was verified by an experiment. In addition, an observation of 160 Gb/s signal by the proposed monitor was demonstrated over 120 minutes using an installed fiber in JGNII testbed.

Synthesis and antiviral activity of 7-benzyl-4-hydroxy-1,5-naphthyridin-2(1H)-one HIV integrase inhibitors.

The medicinal chemistry and structure-activity relationships for a novel series of 7-benzyl-4-hydroxy-1,5-naphthyridin-2(1H)-one HIV-integrase inhibitors are disclosed. Substituent effects were evaluated at the N-1, C-3, and 7-benzyl positions of the naphthyridinone ring system. Low nanomolar IC(50) values were achieved in an HIV-integrase strand transfer assay with both carboxylic ester and carboxamide groups at C-3. More importantly, several carboxamide congeners showed potent antiviral activity in cellular assays. A 7-benzyl substituent was found to be critical for potent enzyme inhibition, and an N-(2-methoxyethyl)carboxamide moiety at C-3 significantly reduced plasma protein binding effects in vitro. Pharmacokinetic data in rats for one carboxamide analogue demonstrated oral bioavailability and reasonable in vivo clearance.

Control and Stabilization of bit-wise phase correlation in 160 (4 x 40) Gbit/s OTDM signal and its impact on transmission.

Detail of control technique of bit-wise phase correlation in 160 (4 x 40) Gbit/s optical time division multiplexing (OTDM) signal using a phase-correlation monitor based on 1-bit delay asymmetric interferometers (AIFs) is described. The 1-bit delay AIF transforms a bit-by-bit optical phase discontinuity to an optical power variation, so that it enables to quantify the phase-jump between adjacent bits. By use of this unique technique, we experimentally demonstrated stable generation of bitwisely phase-controlled 160 Gbit/s periodical alternate-phase return-to-Zero (APRZ) signal in addition to other different modulation formats such as conventional RZ, carrier suppressed RZ (CS-RZ), pair-wise alternate-phase CSRZ (PAP-CSRZ) and pi/2-APRZ. And long term stability was observed with CS-RZ signal. Also, we show some experimental results of 120 km un-repeatered transmission using standard single mode fiber (SSMF) and then discuss the impact of bit-wise phase change on 160 Gbit/s OTDM transmission performance.

Selection of diverse and clinically relevant integrase inhibitor-resistant human immunodeficiency virus type 1 mutants.

Resistance passage studies were conducted with five INIs (integrase inhibitors) that have been tested in clinical trials to date: a new naphthyridinone-type INI S/GSK-364735, raltegravir, elvitegravir, L-870,810 and S-1360. In establishing the passage system and starting from concentrations several fold above the EC(50) value, resistance mutations against S-1360 and related diketoacid-type compounds could be isolated from infected MT-2 cell cultures from day 14 to 28. Q148R and F121Y were the two main pathways of resistance to S/GSK-364735. Q148R/K and N155H, which were found in patients failing raltegravir treatment in Phase IIb studies, were observed during passage with raltegravir with this method. The fold resistance of 40 mutant molecular clones versus wild type virus was compared with these five INIs. The overall resistance pattern of S/GSK-364735 was similar to that of raltegravir and other INIs. However, different fold resistances of particular mutations were noted among different INIs, reflecting a potential to develop INIs with distinctly different resistant profiles.