Status Report 2015 for B–3 Beam Port of KUR
the null alloy.
Furthermore, the data acquisition group of the neutronscience division of KEK (KEK–KENS DAQ group) has
used the B–3 beam port to assess their new 6Li-glass neutron detector system, LiTA12. The LiTA12 system consists of a 6Li-glass neutron detector with a multianodephoto multiplier tube (MA–PMT), an amplifier, and ananalog-to-digital converter (ADC) board. The B–3 beam
port has a wide space around the sample position; therefore we can easily install any other system like the
LiTA12 system.
IMPROVEMENTOF
MONOCHROMATOR
STAGE: The old Cu monochromator stage of the 4CND
was removed (Fig. 2(a)), and then the new Cu monochromator stage was installed at the B–3 beam port (Fig.
2(b)). The Cu monochromator stage consists of two goniometers: rotation and swivel stages (RA07A-W and
SA05B-RM, Kohzu Precision Co., Ltd.).
(b)
4000

1000

PRELIMINARY EXPERIMENTS: The preliminary
neutron diffraction experiments were performed using thehydrogen absorbing alloy, (Ti0.31Cr0.33V0.36)D1.7, where D
is the deuterium. It is worth noting that Ti0.31Cr0.33V0.36 isa null alloy for neutron scattering due to its atomic compositions (i.e., bc[Ti0.31Cr0.33V0.36] = 0; bc[Ti] = –3.370 fm,
bc[Cr] = 3.635 fm, and bc[V] = –0.443 fm), where bc isthe coherent scattering length. The crystal structure has
been refined on the basis of a cubic CaF2-type structurewith a = 4.288(1) Å, using X-ray diffraction with Cu Kα
radiation (see Fig. 1(b)). As shown in Fig. 1(a), we succeeded to observe several Bragg reflections for the(Ti0.31Cr0.33V0.36)D1.7 on the B–3 beam port; which could
be indexed on the basis of λ = 1 Å. Note that the Braggreflections correspond to the D–D correlations because of

70

90

0
FCC

0

INSTRUMENT: The compact multipurpose neutron
diffractometer is now being installed on the B–3 beam
port. The neutron wavelength (λ), which is monochromatized by the (220) plane of a Cu single crystal, is 1 Å. To
cover the detector area (6 º ≤ 2θ ≤ 150 º), 24 3He tube
detectors (1/2 inch in diameter) have been prepared. The
distances from the monochromator to the sample andfrom the sample to the detector will be 1.9 m and 1.2 m,
respectively. A detector bank including the 24 3He tube
detectors will be placed on an arm of the goniometer. In
addition, the new beam shutter was installed at the B-3
beam port.

400

2000

Observed data
Calculated data

110

3000

220

420

Intensity / counts

500

400

1000

Observed data
Calculated data

200

1500

311
222

(a)
220

INTRODUCTION: The B–3 beam port of Kyoto University Research Reactor (KUR) had long been used as afour-circle single-crystal neutron diffractometer (4CND).
For the last decade, however, the 4CND was so old thatits research activity on neutron science was quite low.
Therefore, the 4CND needed to be replaced and a newneutron diffractometer has been required at the B–3 beam
port. Also, the new neutron diffractometer (Compact
multipurpose neutron diffractometer) is critical for the
structural investigations of energy storage materials suchas hydrogen absorbing alloys and rechargeable lithium-ion batteries. The neutron (powder) diffraction is a
powerful tool to determine the positions of light elements(e.g., hydrogen and lithium) in solids. Here, we report thecurrent status of the B-3 beam port of KUR and the preliminary neutron diffraction experiments using a hydrogen absorbing alloy.

222

Research Reactor Institute, Kyoto University (KURRI)
1
High Energy Accelerator Research Organization (KEK)2
Department of Materials and Engineering, Ibaraki University
3
Institute for Materials Research, Tohoku University
4
Graduate School of Engineering, Kyoto University

200

K. Mori, H. Yoshino, Y. Iinuma, T. Fukunaga, Y. Kawabata, S. Sato1, H. Hiraka1, K. Iwase2, Y. Yamaguchi3 andT. Shiono4

Intensity / counts

CO1-1

Laves

20

30

40

50

60

40

70

2θ / °

50

60

80

100

2θ / °

Fig. 1. Neutron and X-ray diffraction data for a hydrogenstorage alloy, (Ti0.31Cr0.33V0.36)D1.7, collected at (a) theB-3 beam port of KUR and (b) a X-ray diffractometerwith Cu Kα radiation.
(a)

(b)

Fig. 2. Improvement of Cu monochromator stage: (a) old
stage, and (b) new stage, respectively.

27011- 61 -

CO1-2

Characterization of Proteolytic Enzymes Derived from NephilaClavata

Y. Hidaka, K. Kouda, Y. Touyama and S. Shimamoto
Graduate School of Science and Engineering Kinki University
INTRODUCTION: Spiders trap insects using a web
net. The fact that they eat them without chewing, indicatethat spiders possesses highly efficient digestive enzymes[1]. Our previous studies suggest that spider’s proteolytic
enzymes are able to digest synthetic spider dragline amyloid fibers [2,3]. Thus, the spider protease has the potential ability to digest amyloid fibers including pathogenic
-amyloid, such as amyloid fibrils, that are associatedwith the development of Alzheimer's disease [1]. Therefore, we purified, identified, and characterized the enzymes derived from Nephila Clavata. The sequence
analyses of the blotted protein on a PVDF membrane
showed N-terminal amino acid residues. Based on thissequence information, we tried to clone the protease
gene.
EXPERIMENTS: The spiders (Nephila Clavata) werecollected in the field of Kyoto University Research Reactor Institute and the spider saliva including proteolytic
enzymes was obtained from Nephila Clavata (50 heads)
by electrical stimulation. The extracts were stored in a
deep freezer until use.
Casein protease assay [4] of the crude spider enzymes
was performed, as previously reported [1]. Peptide substrates for the assay of the protease activity of spider enzymes were chemically synthesized by the Boc solid
phase method, treated with hydrogen fluoride, and purified by reversed phase HPLC [1, 4].
Based on the N-terminal sequence, evidenced by the
Edoman’s degradation method, we prepared generated
primers for the gene cloning and RT-PCR was performed
using mRNA’s from gut cells or full bodies of Nephila
Clavata.
RESULTS: Spider’s digestive fluid that includes
proteolytic enzymes was obtained from Nephila Clavata
by electrical stimulation using a micropipet. The extracts
were applied to SDS-PAGE and the enzymatic activity ofthe protein bands was estimated by a casein protease assay [4]. Two protein bands showed protease activities
and their molecular weights were estimated to be approximately 21.9 and 19.5 kDa, based on the SDS-PAGE
analysis, as we determined previously [1].
To determine the gene sequence of the spider protease,

we tried to obtain its mRNA. For this purpose, generated
primers for the gene cloning of the protease were specifically designed and RT-PCR was performed using mRNA
from gut cells or full bodies of Nephila Clavata. Several
clones were isolated and provided second PCR to produce cDNA’s of spider proteases. The purified candidate
cDNA’s were applied to PCR again, then their cDNA
sequence were directly determined. However, the sequence analysis of the cloned spider protease gene revealed that the cloned genes were non-specifically amplified during RT-PCR. Therefore, generating primers
were re-designed by considering codon bias of spider.
RT-PCR and following PCR produced several candidate
cDNA’s . Their sequence analyses is now in progress.
We also prepared the cDNA of cocoonase to estimate
amyloid digestive activity and expressed it using E. coli
cells. Recombinant cocoonase was successfully
over-expressed as a inclusion body. The protein was purified and analyzed by reversed phase HPLC and
MALD-TOF/MS, respectively. To activate the protein,refolding reactions were carried out under several conditions and its protease activity will be estimated using a
synthetic substrates.
In conclusion, Spider protease was extracted fromits digestive fluid and exhibited strong protease activity.
The protease can be classified as a Ca2+-dependent
carboxypeptidase, based on the results of protease inhibition assays. Spider genes were amplified using generating primers, base on the N-terminal amino acid sequence,by RT-PCR and determined by the direct sequencing
method. However, DNA sequence analysis indicated thatthe cloned DNA was non-specifically amplified during
RT-PCR. The cloning of spider protease genes is in progress. In addition, we also prepared the cDNA of theprotease, cocoonase andover-expressed it using E. coli
expression system. Recombinant cocoonase was successfully refolded as a soluble protein. Its biological activity will be estimated and provide a critical insight tounderstand the amyloid digestive activity of spider andBombyx mori.
REFERENCES:[1] M. Fujiwara et al., Peptide Science, 2013, 447-448(2014).
[2] S. Zheng, et al., Appl. Spectrosc., 43, 1269-1272(1989).
[3] Y. Hidaka, et al., Biopolymers, 96, 222-227 (2011).
[4] C. Dicko, et al., Biochemistry, 43, 1269-1272 (2004).

27040- 62 -

CO1-3Development of a Plano-elliptical Supermirror with Metallic Substrates

M. Hino, T.Hosobata1, N.L.Yamada2, Y.Yamagata1,
S. Takeda3, J. Guo1, S.Morita4, T.Oda, M.Furusaka2 and
Y. Kawabata

lic substrate were conducted at RIKEN [2]. The supermirror coating was conducted with KUR-IBS [3]. The
neutron experiments were conducted at the SOFIA reflectometer at J-PARC/MLF.

Research Reactor Institute, Kyoto Univ., Japan
RAP, RIKEN, Japan
2
IMSS, KEK, Japan
3
Grad. Sch. of Eng., Hokkaido Univ., Japan
4
Tokyo Denki Univ. Japan

1

INTRODUCTION: Progress of neutron optical devices is very significant, however, it is still very difficult forneutron aspherical focusing mirror. The metallic
substrate is robust and ductile, able to withstand highradiation irradiation and high-temperature. It istherefore possible to install them even at a place close
to the neutron target and moderator. Furthermore, it ispossible to fabricate a large focusing supermirror by
mirrorswithcombining
multipleseg-mented
mechanical fastening entailing the usage of screw holes
and fixture tabs. The big problem is required surface
roughness for neutron mirror. It is smaller than 0.5
nm even for m=3 supermirror coating and the mirror
size is large. Here m is the maximum crit-ical angle ofthe mirror in units of critical angle of natural nickel. In
order to realize large ellipsoid neutron focusing
supermirror, we are doing a lot of trials. In this study,
we show the neutron reflectivity of a large planoelliptical supermirror with metallic substrates.
EXPERIMENTS: We fabricated a large plano-elliptical
metallic substrates with electroless nickel-phosphorus(NiP) plating, based on the technology using ultrahighprecision cutting with correction processing, followed bymechanical precision polishing. We manufactured a twosegment large plano-elliptical supermirror with m=3 supermirror coating for the Soft Interface Analyzer (SOFIA) at the Materials and Life Science Experimental Facility
(MLF)
of
Japan
Particle
Accelerator
Research Complex J-PARC [1]. Because the area
limitations of machines used for manufacturing andsupermirror coat-ing. To realize sample focusing optics,
the focusing mir-ror was installed at an equidistantposition between first slit and the sample position. One
focal point was at the first slit. The other was at the
sample position. The major axis of the mirror was
designed to be 2150 mm. The mi-nor axis was designed
to reflect neutrons to use the typi-cal wavelength band (>0.2 nm) for reflectivity measure-ments at SOFIA. For
the mirror coating, the m=3 super-mirror was chosen to
keep as high a reflection angle as possible while
simultaneously keeping the neutron reflec-tivity
sufficiently high. Therefore, the minor axis waschosen as 21.5 mm to reflect the neutrons on the
mirror with an incident angle of 10 mrad. The mirror
length was chosen as 550 mm to avoid interferencewith other de-vices installed to SOFIA. The width was
60 mm, as de-termined from the maximum beam size
width of 50 mm. The manufacturing, polishing andcleaning of the metal-

RESULTS: Figure1 shows neutron reflectivity of the
10 measured locations in the plano-elliptical supermirror
with metallic substrates. Typical surface roughness of the
top surfaces of metallic substrates was approximately 0.3
nm rms using a white light interferometer (NewView
7200; Zygo Corp.).Using the KUR-IBS, m=3 NiC/Ti
supermirrors were coated onto the substrates surface. The
maximum neutron reflectivity of the supermirror on theelliptical metallic substrates was 0.8, which is almost
identical to that with flat glass substrates. However, theaverage of neutron reflectivity was not so high because of
surface contamination resulting from inadequate cleaning.
The figure error shape was 4.6 µm P–V. The slope error
was 33 µrad rms. The focusing supermirror was installedat the SOFIA neutron reflectometer, showing high neutron reflectivity and giving minimal beam width of 0.34
mm in FWHM. Because of the large beam divergence
accepted by the mirror, the count rate with the focusing
mirror was 3.3 times higher than that obtained using
conventional two-slit collimation[2].

Fig. 1. Neutron reflectivity of the 10 measured
locations in the plano-elliptical supermirror withmetallic substrates. The focusing mirror consists of
two segements (Mirror A and B).
REFERENCES:[1] N.L.Yamada, et al.Euro.Phys.J.Plus 126(2011), 108.
[2] S.Takeda, et al., Optics Express (2016), in press..
[3] M.Hino, et al., Nucl. Instr. and Meth., 797(2015) 265.

27043
- 63 -

CO1-4

Neutron Irradiation Effect of High-density MoO3 Pellets for Mo-99 Production (3)T. Ishida, Y. Suzuki, K. Nishikata, M. Yonekawa, Y. Kato,
A. Shibata, A. Kimura, Y. Matsui, K. Tsuchiya,
T. Sano1, Y. Fujihara1 and J. Zhang1
Neutron Irradiation and Testing Reactor Center, JAEA
1
Research Reactor Institute, Kyoto University
INTRODUCTION: JAEA has a plan to produce 99Mo
by (n, γ) method ((n, γ)99Mo production), a parent nuclideof 99mTc. In 2014, irradiation test was carried out withthe high-density molybdenum trioxide (MoO3) pellets in
the hydraulic conveyer (HYD) of the Kyoto University
Research Reactor (KUR) and the 99mTc solution was extracted from the solution of irradiated MoO3) pellets. In
this study, the qualification of 99mTc solution was evaluated.

not detected in the 99mTc solutions of 1st run and 2nd run.
Figure 1 shows the -ray spectrum of Mo solution and99m
Tc solution. MEK content was lower than that of thestandard values (5,000ppm). Osmotic pressure of the99m
Tc solutions was almost the same values of that of thesaline and endotoxin was not detected. From the results,
the high purity 99mTc solutions were obtained by thismethod.
Table 2 Qualification of 99mTc solutions
SampleItems
pH
Radionuclidic
purities
Radiochemical
impurities
MEK content (ppm)

EXPERIMENTS:
The high-density MoO3 pellets
were irradiated in the HYD of the KUR. After the irradiation tests, the irradiated MoO3 pellets were transported
from KUR to JMTR-HL. The irradiated pellets weredissolved with 6M-NaOH solution (Mo Solution) in the
Lead Cell and the 99mTc was extracted from the Mo solution by the solvent extraction method using methyl ethyl
ketone (MEK). Two types of the alumina columns wereprepared for the extraction of 99mTc solution. At first,
MEK containing 99mTc was passed through a basic alumina column for removing impurities such as Mo andH2O. Next, the MEK was flown through the acidic alumina column in order to lead the whole 99mTc. Finally,saline (0.9%-NaCl) was passed the acidic alumina column and 99mTc solution was collected as a product. The
inspection of 99mTc solution was carried out using pH
meter, ICP-AES, HPLC, germanium detector, and so on.

Al content (ppm)
Osmotic pressure(mosm)
Endotoxin

Standard
Values
4.5 – 7.0
≦0.015%

1st run
5.5
99m

Tc(only)

2nd run5-6
99m

Tc(only)

≦5(%)

2.0

-

≦5000

404

-

≦10

-

<5

270 - 300

286

297

N.D.

N.D.

-

RESULTS: After the dissolution of irradiated MoO3
pellets and extraction of 99mTc solution, 99Mo and 99mTc
activities were measured in the Mo solution and 99mTc
solution by the germanium detector. Table 1 shows theresult of 99mTc recovery rates from the Mo solution [1].
The recovery rate of the 1st and 2nd runs achieved the target values (>80%).
Table 1No.
1st run
2nd run99m

Tc recovery rates from the Mo solution
99m

Activity of Tc (Bq)
1.09 × 109
8.53 × 108

Figure 1

Recovery rate (%)
82.3
95.7

Table 2 shows the qualification of 99mTc solutions. In thistable, the standard values were decided under the framework of Strategic Program for Basic Nuclear Researchlaunched by the MEXT. Radionuclidic purities and radiochemical impurities were measured and the values
obtained satisfactory results. Especially, 92mNb observed in the Mo solution as impurities, but 92mNb was

-ray spectrum of Mo and 99mTc solutions.

CONCLUSION: After the irradiation test of thehigh-density MoO3 pellets in the KUR, 99mTc was extracted from the Mo solution. The qualification of 99mTc
solution by the solvent extraction method was satisfied
the standard values for 99mTc radiopharmaceutical products.
REFERENCES:[1] K. Nishikata, et al., Neutron Irradiation Effect ofHigh-density MoO3 pellets for Mo-99 Production (2),
KURR Progress Report 2014, p109.

27066- 64 -

CO1-5Development of High Spatial Resolution Cold / Ultra-cold Neutron Detector

N. Naganawa, S. Awano1, M. Hino2, K. Hirota1,
H. Kawahara1, M. Kitaguchi1, K. Mishima3,and A. Umemoto1

scope with an epi-illumination system.

Institute of Materials and Systems for Sustainability,
Nagoya University
1
Graduate School of Science, Nagoya University
2
Research Reactor Institute, Kyoto University
3
KEK

RESULTS: As shown in Fig. 1, tracks from absorption
events were clearly seen at the depth around the surfaceof the 10B layer.

INTRODUCTION:
We have been developing
cold/ultra-cold neutron detectors with spatial resolutionof several ten nanometers, by using fine-grained nuclear
emulsion [1, 2] with its AgBrI crystal size of 40nm, andnuclides which emit heavily ionizing particles after absorbing neutrons with high absorption cross section. Oneof the aim is to measure the position distribution of ultra-cold neutrons in the quantized states in the earth’s
gravitational field for the test of gravitational inverse-square law and equality of gravitational and inertial mass, as done in previous studies [3, 4], but more
precisely, by 1 or 2 orders of magnitude.
The extremely high resolution of several ten nanometers
should be realized with a 10nm-thick 10B layer, coatedwith nuclear emulsion. When a neutron is absorbed in the10
B layer, an alpha particle and a 7Li will be emitted. After the development, emulsion layer will be observedwith an optical microscope. A track of an alpha particle ora 7Li starting from the absorber layer will be seen in theemulsion layer just next to the absorber layer. To decide
the absorption point, the track will be extrapolated into
the absorber layer. Then, ambiguity of the absorption
position will be limited by the thickness of the absorber.
In this experiment, we made the detector and exposed tocold neutrons to test the detection principle.

EXPERIMENTS: A detector was made in the following procedure. A layer of 10B with thickness of 10nm was
sputtered on a base, a piece of silicon wafer, at KURRI
by M. Hino. The 10B layer was coated with fine-grained
nuclear emulsion melted at the temperature of 40 degrees
centigrade. Thickness of the emulsion layer coated was
10micron. After drying, it was packed in a light-tight bag
made of thin layers of aluminum and polyethylene.
The detector was exposed to cold neutrons with energyof 10meV in average at BL05 in J-PARC MLF. After thedevelopment, observation was done by an optical micro-

Fig.1. Tracks from absorption of neutrons by 10B are
surrounded by red ellipses.
But, we found tracks from absorptions by 10B also inside
the emulsion layer apart from the 10B layer. This showedthat some of 10B moved into emulsion layer. 10B layer
turned out to be instable when it is coated directly by
nuclear emulsion.
We are now developing methods to stabilize it.

REFERENCES:[1] T. Naka and M. Natsume, Nuclear Instruments andMethods A 581 (2007) 761?764[2] T. Naka, T. Asada, et al., Nuclear Instruments andMethods A 718 (2013) 519?521[3] V. V. Nesvizhevsky et al., NATURE |VOL 415 | 17
JANUARY 2002 |.
[4] G. Ichikawa et al., PhysRevLett.112.071101 (2014)

27071- 65 -