Optical loss study of the cryogenic molecular layer using a folded cavity for future gravitational-wave detectors.

In order to increase the number of detectable gravitational-wave sources, future gravitational-wave detectors will operate with cryogenically cooled mirrors. However, recent studies showed that cryogenic mirrors can suffer from the molecular layer formation, which introduces an additional optical loss, and the detector's performance degrades. In order to evaluate the impact of the molecular layer on future cryogenic gravitational-wave detectors, we built a cryogenic folded-cavity setup and developed an ellipsometric measurement method. The optical loss induced by the cryogenic molecular layer shows a large value even at a few nanometer thickness and can deteriorate the performance of the future cryogenic gravitational-wave detectors.

Compact integrated optical sensors and electromagnetic actuators for vibration isolation systems in the gravitational-wave detector KAGRA.

This paper reports on the design and characteristics of a compact module integrating an optical displacement sensor and an electromagnetic actuator for use with vibration-isolation systems installed in KAGRA, the 3-km baseline gravitational-wave detector in Japan. In the technical concept, the module belongs to a family tree of similar modules used in other interferometric gravitational-wave detector projects. After the initial test run of KAGRA in 2016, the sensor part, which is a type of slot sensor, was modified by increasing the spacing of the slot from 5 mm to 15 mm to avoid the risk of mechanical interference with the sensor flag. We confirm that the sensor performance is comparable to that of the previous design despite the modification. We also confirm that the sensor noise is consistent with the theoretical noise budget. The noise level is 0.5 nm/Hz1/2 at 1 Hz and 0.1 nm/Hz1/2 at 10 Hz, and the linear range of the sensor is 0.7 mm or more. We measured the response of the actuator to be 1 N/A and also measured the resistances and inductances of coils of the actuators to confirm consistency with theory. Coupling coefficients among the different degrees of freedom were also measured and shown to be negligible, varying little between designs. A potential concern about thermal noise contribution due to eddy current loss is discussed. As of 2020, 42 of the modules are in operation at the site.

Frequency-Dependent Squeezed Vacuum Source for Broadband Quantum Noise Reduction in Advanced Gravitational-Wave Detectors.

The astrophysical reach of current and future ground-based gravitational-wave detectors is mostly limited by quantum noise, induced by vacuum fluctuations entering the detector output port. The replacement of this ordinary vacuum field with a squeezed vacuum field has proven to be an effective strategy to mitigate such quantum noise and it is currently used in advanced detectors. However, current squeezing cannot improve the noise across the whole spectrum because of the Heisenberg uncertainty principle: when shot noise at high frequencies is reduced, radiation pressure at low frequencies is increased. A broadband quantum noise reduction is possible by using a more complex squeezing source, obtained by reflecting the squeezed vacuum off a Fabry-Perot cavity, known as filter cavity. Here we report the first demonstration of a frequency-dependent squeezed vacuum source able to reduce quantum noise of advanced gravitational-wave detectors in their whole observation bandwidth. The experiment uses a suspended 300-m-long filter cavity, similar to the one planned for KAGRA, Advanced Virgo, and Advanced LIGO, and capable of inducing a rotation of the squeezing ellipse below 100 Hz.

Measuring scattering light distributions on high-absorptive surfaces for stray-light reduction in gravitational-wave detectors.

In order to establish scattering measurements in material investigations for gravitational-wave detectors, we have built-up devices for measuring the hemispherical scattering distribution of materials which are planned to be used in those detectors as suppressors of scattered light. The measurement benches we have built, a hemispherical goniometer and a direct back-scatterometer, have a maximum background noise of ∼10 -4sr -1 BRDF at 1.064 µm wavelength which is the wavelength of the laser-light for our large interferometer for detecting gravitational waves, KAGRA. With these instruments, we have characterized the surface scattering of, e.g., NiP platings, metals, and different carbonaceous coatings, which are supposed to minimize the amount of scattered light in interferometers. The three most important materials for KAGRA's construction (SiC, "Solblack", and "VantaBlack") are presented in this paper. Furthermore, we will try to explain the scattering distributions with the generalized Harvey-Shack model (smooth-surface approximation) which is a common method for surface-scattering calculations. At the end, we give also some valuations about the vacuum compatibility of the materials, which is important for instruments like KAGRA that work under ultra-high vacuum conditions.

Demonstration of Displacement Sensing of a mg-Scale Pendulum for mm- and mg-Scale Gravity Measurements.

Gravity generated by large masses has been observed using a variety of probes from atomic interferometers to torsional balances. However, gravitational coupling between small masses has never been observed so far. Here, we demonstrate sensitive displacement sensing of the Brownian motion of an optically trapped 7 mg pendulum motion whose natural quality factor is increased to 10^{8} through dissipation dilution. The sensitivity for an integration time of one second corresponds to the displacement generated in a millimeter-scale gravitational experiment between the probe and a 100 mg source mass, whose position is modulated at the pendulum mechanical resonant frequency. Development of such a sensitive displacement sensor using a milligram-scale device will pave the way for a new class of experiments where gravitational coupling between small masses in quantum regimes can be achieved.

Calculation method for light scattering caused by multilayer coated mirrors in gravitational wave detectors.

Scattered light in inteferometric gravitational wave detectors needs to be reduced so that it will not harm the actual signals coming from a gravitational wave. In this paper, we report on the application of the theory of light scattering from mirrors in interferometric detectors having multilayer coatings on their surfaces and compared the results with single-surface scattering theories, which are traditionally used in the field of gravitational wave detectors. For the first time in this field, we have calculated the scattering distributions of the power-recycling, the signal-recycling, and the beam-splitter mirrors in KAGRA (a cryogenic interferometric gravitational wave detector currently under construction in the Kamioka mine in Japan) by using models of their multilayer coatings. Furthermore, we have performed simulations to show the differences between multilayer scattering and single-surface scattering models in the back-scattering of mechanical structures close to the mirrors and the impact on the sensitivity of the KAGRA detector. We show that the back-scattering by using those coatings can be larger by up to almost two orders of magnitude and they also give rise to additional scattering features that should be taken into account for all optical applications in gravitational wave detectors.

Characterization of the room temperature payload prototype for the cryogenic interferometric gravitational wave detector KAGRA.

KAGRA is a cryogenic interferometric gravitational wave detector currently under construction in the Kamioka mine in Japan. Besides the cryogenic test masses, KAGRA will also rely on room temperature optics which will hang at the bottom of vibration isolation chains. The payload of each chain comprises an optic, a system to align it, and an active feedback system to damp the resonant motion of the suspension itself. This article describes the performance of a payload prototype that was assembled and tested in vacuum at the TAMA300 site at the NAOJ in Mitaka, Tokyo. We describe the mechanical components of the payload prototype and their functionality. A description of the active components of the feedback system and their capabilities is also given. The performance of the active system is illustrated by measuring the quality factors of some of the resonances of the suspension. Finally, the alignment capabilities offered by the payload are reported.

Purification and characterization of cocoonase from the silkworm Bombyx mori.

Cocoonase (CCN) which facilitates the degradation of a cocoon is recognized as a trypsin-like serine protease. In this study, CCN from the silkworm Bombyx mori was purified and comprehensively characterized. Its activity was maximal at about pH 9.8. It was stable above pH 3.4 at 4 °C and below 50 °C at pH 7.5. CuSO4, FeSO4, and ZnSO4 showed inhibitory effects on CCN, but other salts improved activity. Typical trypsin inhibitors inhibited CCN, but the relative inhibitory activities were much lower than those against bovine trypsin. An extract of cocoon shells inhibited trypsin, but it was only slightly inhibitory against CCN. There were significant differences in catalytic efficiencies and substrate specificities as between CCN and bovine trypsin.

Optically trapped mirror for reaching the standard quantum limit.

The preparation of a mechanical oscillator driven by quantum back-action is a fundamental requirement to reach the standard quantum limit (SQL) for force measurement, in optomechanical systems. However, thermal fluctuating force generally dominates a disturbance on the oscillator. In the macroscopic scale, an optical linear cavity including a suspended mirror has been used for the weak force measurement, such as gravitational-wave detectors. This configuration has the advantages of reducing the dissipation of the pendulum (i.e., suspension thermal noise) due to a gravitational dilution by using a thin wire, and of increasing the circulating laser power. However, the use of the thin wire is weak for an optical torsional anti-spring effect in the cavity, due to the low mechanical restoring force of the wire. Thus, there is the trade-off between the stability of the system and the sensitivity. Here, we describe using a triangular optical cavity to overcome this limitation for reaching the SQL. The triangular cavity can provide a sensitive and stable system, because it can optically trap the mirror's motion of the yaw, through an optical positive torsional spring effect. To show this, we demonstrate a measurement of the torsional spring effect caused by radiation pressure forces.

Salt-induced changes in the subunit structure of the Bacillus stearothermophilus lipoate acetyltransferase.

The Bacillus stearothermophilus lipoate acetyltransferase (E2), composed of sixty identical, subunits is the core component of the pyruvate dehydrogenase complex (PDC). E2 polypeptide is composed of LD, PSBD, and CD domains. Most studies had focused on a truncated E2 that is deficient in LD and PSBD, because CD mainly contributes to maintaining the multimeric structure. We examined salt-induced changes in E2 without truncation and constructed reaction models. We speculate that in the presence of KCl, E2 is dissociated into a monomer and then assembled into an aggregative complex (C(A)) and a quasi-stable complex (C(Q)). C(A) was larger than C(Q), but smaller than intact E2. C(A) and C(Q), were dominant complexes at about neutral pH and at basic pH respectively. PDC, in which PSBD is occupied by other components, and a truncated E2 undergo dissociation only. LD-PSBD region besides CD might then contribute to the partial association of dissociated E2.

New limit on Lorentz violation using a double-pass optical ring cavity.

A search for Lorentz violation in electrodynamics was performed by measuring the resonant frequency difference between two counterpropagating directions of an optical ring cavity. Our cavity contains a dielectric element, which makes our cavity sensitive to the violation. The laser frequency is stabilized to the counterclockwise resonance of the cavity, and the transmitted light is reflected back into the cavity for resonant frequency comparison with the clockwise resonance. This double-pass configuration enables a null experiment and gives high common mode rejection of environmental disturbances. We found no evidence for odd-parity anisotropy at the level of δc/c ≲ 10(-14). Within the framework of the standard model extension, our result put more than 5 times better limits on three odd-parity parameters κ(o+)(JK) and a 12 times better limit on the scalar parameter κ(tr) compared with the previous best limits.

Structural basis for catalytic activity of a silkworm Delta-class glutathione transferase.

BACKGROUND: Glutathione transferase (GST) catalyzes glutathione conjugation, a major detoxification pathway for xenobiotics and endogenous substances. Here, we determined the crystal structure of a Delta-class GST from Bombyx mori (bmGSTD) to examine its catalytic residues. METHODS: The three-dimensional structure of bmGSTD was resolved by the molecular replacement method and refined to a resolution of 2.0Å. RESULTS: Structural alignment with a Delta-class GST of Anopheles gambiae indicated that bmGSTD contains 2 distinct domains (an N-terminal domain and a C-terminal domain) connected by a linker. The bound glutathione localized at the N-terminal domain. Putative catalytic residues were changed to alanine by site-directed mutagenesis, and the resulting mutants were characterized in terms of catalytic activity using glutathione and 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTD mutants indicated that Ser11, Gln51, His52, Ser67, and Arg68 are important for enzyme function. GENERAL SIGNIFICANCE: These results provide structural insights into the catalysis of glutathione conjugation in B. mori by bmGSTD.

Crystallographic survey of active sites of an unclassified glutathione transferase from Bombyx mori.

BACKGROUND: Glutathione transferase (GST) catalyzes a major step in the xenobiotic detoxification pathway. We previously identified a novel, unclassified GST that is upregulated in an insecticide-resistant silkworm (Bombyx mori) upon insecticide exposure. Here, we sought to further characterize this GST, bmGSTu, by solving and refining its crystal structure and identifying its catalytic residues. METHODS: The structure of wild-type bmGSTu was determined with a resolution of 2.1Å by synchrotron radiation and molecular modeling. Potential catalytic residues were mutated to alanine by means of site-directed mutagenesis, and kinetic data determined for wild-type and mutated bmGSTu. RESULTS: We found that bmGSTu occurred as a dimer, and that, like other GSTs, each subunit displayed a G-site and an H-site in the active center. Bound glutathione could be localized at the G-site. Kinetic data of the mutated forms of bmGSTu show that Val55, Glu67, and Ser68 in the G-site are important for catalysis. Furthermore, the H-site showed some unique features. CONCLUSIONS: This is the first study to our knowledge to elucidate the molecular conformation of this B. mori GST. Our results indicate that residues Val55, Glu67, and Ser68, as well as Tyr7 and Ser12, in the glutathione-binding region of bmGSTu are critical for catalytic function. GENERAL SIGNIFICANCE: Our results, together with our previous finding that bmGSTu was preferentially induced in an insecticide-resistant strain, support the idea that bmGSTu functions in the transformation of exogenous chemical agents. Furthermore, the unique features observed in bmGSTu may shed light on mechanisms of insecticide resistance.

Upper limit on gravitational wave backgrounds at 0.2 Hz with a torsion-bar antenna.

We present the first upper limit on gravitational wave (GW) backgrounds at an unexplored frequency of 0.2 Hz using a torsion-bar antenna (TOBA). A TOBA was proposed to search for low-frequency GWs. We have developed a small-scaled TOBA and successfully found Ω(gw)(f)<4.3×10(17) at 0.2 Hz as demonstration of the TOBA's capabilities, where Ω(gw)(f) is the GW energy density per logarithmic frequency interval in units of the closure density. Our result is the first nonintegrated limit to bridge the gap between the LIGO band (around 100 Hz) and the Cassini band (10(-6)-10(-4) Hz).

Molecular characterization of an insecticide-induced novel glutathione transferase in silkworm.

BACKGROUND: The glutathione transferase (GST) superfamily is involved in the detoxification of various xenobiotics. We have identified a GST mRNA that was induced in the fat bodies of a silkworm strain exhibiting diazinon resistance and have investigated the enzyme properties of this GST. METHODS: A soluble recombinant protein was overexpressed in Escherichia coli. Amino acid residues of interest were changed to alanine by site-directed mutagenesis. RESULTS AND CONCLUSIONS: Phylogenetic analysis of the deduced amino acid sequence indicates that this GST belongs to an unclassified group previously reported in mosquitoes. This enzyme, named bmGSTu, has highly conserved amino acid residues, including Tyr7, Ser12 and Asn50. A recombinant bmGSTu was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTu mutants indicated that Tyr7, Ser12 and Asn50 are involved in enzyme function. GENERAL SIGNIFICANCE: These results support the hypothesis that bmGSTu may play a role in insecticide resistance in Bombyx mori.

Identification and expression analysis of minichromosome maintenance proteins in the silkworm, Bombyx mori.

The minichromosome maintenance protein (MCM) family is involved in the regulatory role of DNA replication in eukaryotic organisms. A cDNA encoding of an MCM of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequenced. The resultant amino acid sequence and phylogenetic analysis revealed high identity to MCM, and specifically to MCM7, of vertebrates and invertebrates. An RT-PCR showed that the bmMCM7 transcript was present in the ovaries, testes, silk glands, and fat bodies of larval silkworms. Expression plasmids were transformed into competent Escherichia coli and overexpressed. This is the first report on the identification of MCM helicase of the silkworm, B. mori.

Structural properties of silkworm small heat-shock proteins: sHSP19.9 and sHSP20.8.

sHSP20.8 and sHSP19.9 are silkworm small-heat shock proteins (sHSPs) comprising a number of polypeptides of molecular sizes of several tens of kilodaltons as subunits. The structural properties of sHSPs were investigated. sHSP19.9 was found to be aggregated by itself during incubation at 60 degrees C. Aggregation was suppressed in the presence of dithiothreitol and at high ionic strength. In contrast, sHSP20.8 was not aggregated. Aggregation of sHSP19.9 was partially suppressed by sHSP20.8 and in the presence of catalase as a target protein. Based on changes in small-angle X-ray scattering, it is possible that the molecular size of sHSP19.9 is larger than that of sHSP20.8, and that their molecular sizes increase with increasing temperature in a reversible, biphasic manner. sHSPs did not protect catalase from thermal inactivation, but protected it from precipitation by forming a soluble complex. sHSP20.8 and sHSP19.9 with dithiothreitol were stable against lyophilization, autoclaving at 120 degrees C, and boiling.

Biochemical properties of an omega-class glutathione S-transferase of the silkmoth, Bombyx mori.

A cDNA encoding an omega-class glutathione S-transferase of the silkmoth, Bombyx mori (bmGSTO), was cloned by reverse transcriptase-polymerase chain reaction. The resulting clone was sequenced and deduced for amino acid sequence, which revealed 40, 40, and 39% identities to omega-class GSTs from human, pig, and mouse, respectively. A recombinant protein (rbmGSTO) was functionally overexpressed in Escherichia coli cells in a soluble form and purified to homogeneity. rbmGSTO was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GST, as well as with 4-hydroxynonenal, a product of lipid peroxidation. This enzyme was shown to have high affinity for organophosphorus insecticide and was present abundantly in silkmoth strain exhibiting fenitrothion resistance. These results indicate that bmGSTO could be involved in the increase in level of insecticide resistance for lepidopteran insects.

Proteomic profiling of the silkworm skeletal muscle proteins during larval-pupal metamorphosis.

The silkworm is a typical holometabolous insect going through drastic morphological changes upon metamorphosis from larvae to pupae. Comprehensive studies focusing on the changes help elucidate understanding of a biogenic mechanism. Here, we report the initial profile of the intersegmental muscle (ISM) proteins of the silkworm during larval-pupal metamorphosis. In total, 258 protein spots were resolved by two-dimensional gel electrophoresis (2-DE). Fifty-seven larval proteins were identified, where 3 proteins were exclusively detected in larval samples. Fifty-four other proteins were common in pupal samples. Of these, 12 proteins belonging to the contractile apparatus, metabolism, regulation, and signal transduction were altered in their contents during the metamorphosis from larvae to pupae. Three pupa-defective proteins were identified as isoforms of troponin I, followed by an immunoblotting validation. This data will be helpful in understanding the biochemistry of an insect ISM.

Expression and characterization of a sigma-class glutathione S-transferase of the fall webworm, Hyphantria cunea.

A cDNA encoding glutathione S-transferase (GST) of the fall webworm, Hyphantria cunea, was cloned by reverse transcriptase-polymerase chain reaction. The resulting clone (hcGST) was sequenced and deduced for amino acid sequence, which revealed 87, 59, and 42% identities to Sigma-class GSTs from Bombyx mori, Manduca sexta, and Blattella germanica respectively. A recombinant hcGST protein (rhcGST) was functionally overexpressed in Escherichia coli cells in a soluble form and purified to homogeneity. rhcGST retained more than 75% of its original GST activity after incubation at pHs 6 to 11. Incubation for 30 min at temperatures below 50 degrees C scarcely affected the activity. rhcGST was able to catalyze the reaction of glutathione with 1-chloro-2,4-dinitrobenzene, a universal substrate for GST, as well as with 4-hydroxynonenal, a product of lipid peroxidation. We also found that as compared to B. mori Sigma-class GST, rhcGST had a higher affinity for fenitrothion, an organophosphorus insecticide.