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We developed a CANDLES-III system to study the neutrino-less double beta (0$\nu\beta\beta$) decay of $^{48}$Ca. The proposed system employs 96 CaF$_{2}$ scintillation crystals (305 kg) with natural Ca ($^{\rm nat.}$Ca) isotope which corresponds 350\,g of $^{48}$Ca. External backgrounds were rejected using a 4$\pi$ active shield of a liquid scintillator surrounding the CaF$_2$ crystals. The internal backgrounds caused by the radioactive impurities within the CaF$_2$ crystals can be reduced effectively through analysis of the signal pulse shape. We analyzed the data obtained in the Kamioka underground for a live-time of 130.4\,days to evaluate the feasibility of the low background measurement with the CANDLES-III detector. Using Monte Carlo simulations, we estimated the background rate from the radioactive impurities in the CaF$_{2}$ crystals and the rate of high energy $\gamma$-rays caused by the (n, $\gamma$) reactions induced by environmental neutrons. The expected background rate was in a good agreement with the measured rate, i.e., approximately 10$^{-3}$ events/keV/yr/(kg of $^{\rm nat.}$Ca), in the 0$\nu\beta\beta$ window. In conclusion, the background candidates were estimated properly by comparing the measured energy spectrum with the background simulations. With this measurement method, we performed the first search for 0$\nu\beta\beta$ decay in a low background condition using a detector with a Ca isotope, in which the Ca present was not enriched, in a scale of hundreds of kg. The $^{48}$Ca isotope has a high potential for use in 0$\nu\beta\beta$ decay search, and is expected to be useful for the development of a next-generation detector for highly sensitive measurements.

High energy γ rays with several MeV produced by (n,γ) reactions can be a trouble for low background measurements in the underground laboratories such as double beta decay experiments. In the CANDLES project, which aimed to observe the neutrino-less double beta decay from 48Ca, γ rays caused by (n,γ) reactions were found to be the most significant background. The profile of the background was studied by measurements with a neutron source and a simulation with a validity check of neutron processes in Geant4. The observed spectrum of γ rays from (n,γ) reactions was well reproduced by the simulated spectra, which were originated from the surrounding rock and a detector tank made of stainless steel. The environmental neutron flux was derived by the observed event rate of γ rays from (n,γ) reactions using the simulation. The thermal and non-thermal neutron flux were found to be ( 1.3 ± 0.6 ) × 10 − 6 cm − 2 s − 1 and ( 1.1 ± 0.5 ) × 10 − 5 cm − 2 s − 1 , respectively. It is necessary to install an additional shield to reduce the background from (n,γ) reaction to the required level.

CAlcium fluoride for the study of Neutrinos and Dark matters by Low-energy Spectrometer (CANDLES) searches for neutrino-less double beta decay of $^{48}$Ca using a CaF$_2$ scintillator array. A high Q-value of $^{48}$Ca at 4,272 keV enabled us to achieve very low background condition, however, at the same it causes difficulties in calibrating the detector's Q-value region because of the absence of a standard high-energy $\gamma$-ray source. Therefore, we have developed a novel calibration system based on $\gamma$-ray emission by neutron capture on $^{28}$Si, $^{56}$Fe and $^{58}$Ni nuclei. In the paper, we report the development of the new calibration system as well as the results of energy calibration in CANDLES up to 9 MeV., Comment: 6 pages, 7 figures

The neutrino-less double beta decay (0νββ) is a powerful tool to probe the neutrino mass. The 0νββ is forbidden in Standard Model (SM) due to the violation of lepton number conservation. In experiments searching for 0νββ, the two-neutrino double beta decay (2νββ), which is an allowed process in SM, behaves as an irreducible background within the energy of interest. To discriminate the 0νββ from 2νββ, the energy resolution must be improved. The CANDLES experiment is searching for the 0νββ of 48Ca using CaF 2 (pure) scintillator as the detector and the source. Scintillation photons are collected by Photo Multiplier Tubes (PMTs) surrounding the crystals. A Flash Analog-to-Digital Converter records the waveform of each PMT. Because of the long decay time of CaF 2 (1 µsec), we make a signal integral of 4 µsec to calculate the obtained energy. Thus, the other fluctuation in the baseline is accumulated in the signal integration, and it makes the energy resolution worse. To reduce this fluctuation, we can count the number of photoelectrons in every PMT. Still, we have a significant overlapping probability of single photoelectron signals. This overlapping probability implies the photoelectrons lost, which causes terrible energy resolution. To improve the energy resolution, we introduce an alternative method named "partial photon counting" by dividing the waveform into two parts, then, perform signal integral in the prompt part and photon counting the second part. As a result, the energy resolution is reduced 4.0-3.5 % at 1460 keV (γ-ray of 40K) and 3-2.7 % at 2615 keV (γ-ray of 208Tl).

Filial imprinting in precocial birds is a useful model for studying early learning and cognitive development, as it is characterized by a well-defined sensitive or critical period. We recently showed that the thyroid hormone 3,5,3′-triiodothyronine (T3) determines the onset of the sensitive period. Moreover, exogenous injection of T3 into the intermediate medial mesopallium (IMM) region (analogous to the associative cortex in mammals) enables imprinting even on post-hatch day 4 or 6 when the sensitive period has been terminated. However, the neural mechanisms downstream from T3 action in the IMM region remain elusive. Here, we analyzed the functional involvement of the intermediate hyperpallium apicale (IMHA) in T3 action. Bilateral excitotoxic ablation of the IMHA prevented imprinting in newly hatched chicks, and also suppressed the recovery of the sensitive period by systemic intra-venous or localized intra-IMM injection of T3 in day-4 chicks. In contrast to the effect in the IMM, direct injection of T3 into the IMHA did not enable imprinting in day-4 chicks. Moreover, bilateral ablation of IMHA after imprinting training impaired recall. These results suggest that the IMHA is critical for memory acquisition downstream following T3 action in the IMM and further, that it receives and retains information stored in the IMM for recall. Furthermore, both an avian adeno-associated viral construct containing an anterograde tracer (wheat-germ agglutinin) and a retrograde tracer (cholera toxin subunit B) revealed neural connections from the IMM to the IMHA. Taken together, our findings suggest that hierarchical processes from the primary area (IMM) to the secondary area (IMHA) are required for imprinting.

CANDLES aims to measure the very rare neutrino-less double beta decays using 48Ca which has the highest value among all isotope candidates. Low background techniques are the key to observe such rare phenomena. The largest background was found to be the high energy gamma rays from neutron captures on abundant material near the detector. The shielding system consisted of lead and B-contained rubber sheet was constructed to reduce the background. Performance of the constructed shielding system were described in this paper.

The neutrino-less double beta decay is acquiring great interest after the confirmation of neutrino oscillation which demonstrated nonzero neutrino mass. In order to search for neutrino-less double beta decay of $^{48}$Ca, we proposed the CANDLES project by using CaF$_{2}$(pure) scintillators. Now we installed the CANDLES III system at the Kamioka underground laboratory. The CANDLES III system realizes the low background condition by a characteristic structure and data analyses for background rejection. Furthermore we installed new shielding system in order to reduce $\gamma$-ray backgrounds from neutron capture reaction. Here we report performances of the CANDLES III system.

We have studied the neutrino-less double beta decay(0νββ) of 48Ca by using CaF2(pure) scintillators. Analysis for rejection of backgrounds(212Bi→ 212Po events and 208Tl events) was effective to reduce backgrounds in Qββ-value region. No events are observed in the Qββ-value region for the data of 131 days × 86 kg. It gives a lower limit (90% confidence level) of (math) > 6.2 × 1022 year (preliminary) for the half-life of 0νββ of 48Ca.

Made available in DSpace on 2018-12-11T16:40:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Hookworm infection stands out for its worldwide distribution and for its veterinary and public health relevance. Based on copromicroscopic examinations and polymerase chain reaction (PCR) amplification of the ITS1-5.8S-ITS2 region, we assessed, respectively, the prevalence of intestinal parasites and the identification of canine hookworm species in faeces recovered from 278 dogs living in households of an inland municipality of São Paulo State, Brazil. Intestinal parasites were found in 67.3% of dogs and hookworm infection was found at the highest prevalence rate (56.6%), followed by Toxocara canis (11.9%), Isospora spp. (11.9%), Giardia spp. (5.8%), Sarcocystis spp. (4.0%), 'Hammondia-like' (1.4%), Dipylidium caninum (1.1%) and Trichuris vulpis (0.7%). Of 158 samples positive for hookworm eggs, 106 (67.1%) were amplified by PCR and, of those, 88 (55.7%) were successfully sequenced for species identification. Single infections with Ancylostoma caninum and Ancylostoma braziliense were recorded in 61.4% and 12.5%, respectively, and mixed infections were found in 26.1%. The nucleotide sequences of both species showed high identity rates (98-100%) when compared with reference sequences. Although A. caninum was the most prevalent hookworm in the dogs assessed, the occurrence of both A. caninum and A. braziliense in single and/or mixed infections poses a potential risk for the local population in a low-income area, especially children, to acquire cutaneous larva migrans (CLM). UNESP - Universidade Estadual Paulista Departamento de Parasitologia Instituto de Biociências UFS - Universidade Federal de Sergipe Departamento de Morfologia Centro de Ciências Biológicas e da Saúde USC - Universidade Do Sagrado Coração Departamento de Ciências Biológicas e da Saúde UNESP - Universidade Estadual Paulista Departamento de Parasitologia Instituto de Biociências FAPESP: 06/56151-3

Coprological examination was used to estimate the prevalence of gastrointestinal parasites in stray and domiciled dogs from Botucatu, Sao Paulo State, Brazil. Risk factors for dog infection were assessed in relation to demographic, husbandry and management data. The dog owners completed a questionnaire survey on some aspects of dog parasitism such as parasite species, mechanisms of infection, awareness of zoonotic diseases and history of anthelmintic usage. Parasites were found in the faeces of 138 dogs, with an overall prevalence of 54.3%. Dogs harbouring one parasite were more common (31.4%) than those harbouring two (18.5%), three (3.2%) or four (1.2%). The following parasites and their respective frequencies were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Dipylidium caninum (2.4%), Isospora (3.5%), Cryptosporidium (3.1%) and Sarcocystis (2.7%). Stray dogs were found more likely to be poliparasitized (P 0.05). Except for Ancylostoma, that showed a significantly higher prevalence in dogs living in a multi-dog household (P

Partial resistance to rice blast in the Oryza sativa japonica group cv. Chubu 32 is controlled by Pi34, a major quantitative trait locus (QTL) on chromosome 11, and several uncharacterized QTLs. The objectives of the study were (i) high-resolution genetic and physical mapping of Pi34 and (ii) identification of new QTL imparting resistance to rice blast. Chubu 32 was crossed with a susceptible chromosomal segment substitution line (CSSL) of cv. Koshihikari. From 4,012 of segregating individuals, 213 recombinants in the Pi34 region were screened by using polymerase chain reaction-based markers and tested resistance in the field and greenhouse. The Pi34 locus is located in the 54.1-kb region on the genomic sequence of cv. Nipponbare. We constructed a bacterial artificial chromosome (BAC) library of Chubu 32, selected the clone containing Pi34, and sequenced it. The Pi34 locus consequently was located on an interval of 65.3 kb containing 10 predicted open reading frames (ORFs). Two of these ORFs were predicted only in Chubu 32 and encoded transposable elements. The other eight ORFs were found in both Chubu 32 and Nipponbare and one of them, which encoded an unknown protein, showed significantly different amino acid sequences between two cultivars. The new QTL, Piq6(t), was detected on the short arm of chromosome 6 and the genetic distance of flanking markers was 16.9 centimorgans in Nipponbare. Pi34 and Piq6(t) acted additively on resistance to rice blast but the effect of Piq6(t) was relatively small compared with Pi34.

SUMMARY 1. The aim of the present study was to examine the effects of moderate exercise on serum lipids, lipoproteins and apolipoproteins in seven sedentary young women under controlled conditions. 2. The subjects exercised on separate days for 30 or 60 min at an intensity of 60% of maximal oxygen uptake on a cycle ergometer. Total cholesterol, triglycerides, high-density lipoprotein–cholesterol (HDL-C), HDL2-C, HDL3-C, low-density lipoprotein–cholesterol, apolipoproteins A-I, A-II and B were measured in the serum at the end of the 60 min rest period before each exercise, immediately after the performance of each exercise and at 30 min and 1, 2 and 24 h after each exercise. 3. The results showed that there were no significant differences between the pre- and postexercise samples for any of the parameters tested. 4. The results of the present study suggest that a single bout of exercise designed to simulate a typical training workout has no noticeable effect on serum lipids, lipoproteins and apolipoproteins in normal sedentary young women who have normal lipid profiles, are in the follicular phase of their menstrual cycle and who consume a relatively low-fat diet.

Coal combustion with O2/CO2 is promising because of its easy CO2 recovery, extremely low NOx emission and high sulfation efficiency. The mechanism of direct sulfation of limestone under high CO2 concentration, different from that of CaO–SO2 sulfation, is one of the factors to account for its high sulfation efficiency. However, many unknowns about the mechanism of direct sulfation exist. The mechanisms and kinetics of direct sulfation were examined. Both experiment and modeling suggested that sintering is much mitigated during direct sulfation of limestone. The rate of direct sulfation does not decrease so much with sulfation degree as CaO–SO2 sulfation. Direct sulfation is favorable for high sulfation degree. Furthermore, the diffusivity in the product layer demonstrates high temperature dependence and hardly changes with sulfation degree. This result is associated with the fact that the calcium sulfate layer produced is porous owing to CO2 formation.

The period length of the circadian conidiation rhythm was examined in a mutant strain of Neurospora crassa, un-18, that is temperature sensitive for mycelial growth. The un-18 mutant showed a temperature-sensitive phenotype with respect to both mycelial growth and the period length of the conidiation rhythm. Below 22 degrees C, the un-18 mutation did not affect the period length, but at temperatures between 22 degrees C and 32 degrees C, the period length of the un-18 mutant was approximately 2 h longer than that of the wild-type strain. The un-18+ gene was cloned and was found to encode the second-largest subunit of RNA polymerase I, which is involved in the synthesis of rRNA. These results indicate that a defect in ribosome synthesis, which must result in a lower rate of protein synthesis, lengthens the period of the circadian conidiation rhythm in Neurospora.

AIM: To determine whether transforming growth factor-beta 1 (TGF-beta 1) has a pathogenetic role in disease of the salivary glands. METHODS: An indirect immunohistochemical technique was used to analyse TGF-beta 1 expression in six specimens of normal salivary gland and 23 surgical specimens. RESULTS: TGF-beta 1 was strongly expressed in the ductal epithelial cells of normal salivary gland tissues (six of six cases) and in inflammatory conditions (eight of 11 cases). In contrast, TGF-beta 1 was not detectable in ductal epithelial cells expressing HLA-DR around infiltrating CD4+ CD45RO+ activated T cells, in the salivary gland tissue of patients with Sjögren's syndrome. CONCLUSION: Because TGF-beta 1 has an essential role in the mucosal immunity of salivary glands, abnormal expression of this cytokine must be regarded as a candidate in the pathogenesis of Sjögren's syndrome.

To better understand genome structure and the expression of α/β-gliadin multigenes in hexaploid wheat, bacterial artificial chromosome (BAC) clones containing α/β-gliadin genes from the three loci, Gli-A2, Gli-B2, and Gli-D2, were screened. Based on their restriction fragment patterns, we selected five BAC clones, namely, two clones for Gli-A2, two clones for Gli-B2, and one clone for Gli-D2, to fully sequence. Approximately 200 kb was sequenced for each locus. In total, twelve α/β-gliadin intact genes and four pseudogenes were found, and retrotransposons or other transposons existed in each BAC clone. Dot-plot analysis revealed the pattern of genome segmental duplication within each BAC. We calculated time since duplication of each set of α/β-gliadin genes and insertion of retrotransposons. Duplication of all adjacent genes within the same BAC clone took place before or after allotetrapolyploidization, but duplication of certain genes occurred before diploid differentiation of wheat species. Retrotransposons were also inserted before and after the segmental duplication events. Furthermore, translocation of α/β-gliadin genes from chromosomes 1 to 6 apparently occurred before the diversification of various wheat genomes. Duplication of genome segments containing α/β-gliadin genes and retrotransposons were brought about through unequal crossing-over or saltatory replication and α/β-gliadin genes per se were duplicated without any recombination events. Out of twelve intact α/β-gliadin genes detected from their sequences, nine were expressed, although their patterns of expression were distinct. Since they have similar cis-elements and promoter structures, the mechanisms underlying their distinct gene expression and possible applications are discussed.

Hepatocellular carcinoma (HCC) shares only a small proportion of hyperechoic liver tumors detected in MHTS and mimics hemangioma on ultrasonography (US). The precise diagnosis of HCC is hence difficult in MHTS, but presence or absence of underlying chronic liver disease and serum alfa-fetoprotein (AFP) titer might provide useful informations.

Airborne grass pollen in a grassland was measured using a Burkard air sampler and immunoblotting. The pollen antigens collected on the tape of the air sampler were transferred onto a nitrocellulose membrane, and visualized as purple spots by immunoenzymatic staining with anti-Lol p I antibody. These spots were quantified automatically using an image processing system. The airborne grass pollen antigens in the grassland where four species of grass were growing were prevalent mainly from evening to early morning during the first half of the pollen season, but in the daytime during the latter half of the season. The antigen dispersion time was confined to a few hours of the day. The flowering time of several grass species which were hydroponically cultured in the laboratory were also studied. Each species had a different flowering time. We believe that the time when an antigen is present during the day is related to the different flowering times of the various grass species growing in the grassland.

Coprological examination was used to estimate the prevalence of gastrointestinal parasites in stray and domiciled dogs from Botucatu, São Paulo State, Brazil. Risk factors for dog infection were assessed in relation to demographic, husbandry and management data. The dog owners completed a questionnaire survey on some aspects of dog parasitism such as parasite species, mechanisms of infection, awareness of zoonotic diseases and history of anthelmintic usage. Parasites were found in the faeces of 138 dogs, with an overall prevalence of 54.3%. Dogs harbouring one parasite were more common (31.4%) than those harbouring two (18.5%), three (3.2%) or four (1.2%). The following parasites and their respective frequencies were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Dipylidium caninum (2.4%), Isospora (3.5%), Cryptosporidium (3.1%) and Sarcocystis (2.7%). Stray dogs were found more likely to be poliparasitized (P0.01) and presented higher prevalence of Ancylostoma, T. canis and Giardia (P0.01) than domiciled ones. Toxocara canis was detected more frequently in dogs with6 months of age (P0.05) and no effect of sex or breed could be observed (P0.05). Except for Ancylostoma, that showed a significantly higher prevalence in dogs living in a multi-dog household (P0.01), parasite prevalences were similar in single- and multi-dog household. The answers of dog owners to the questionnaire showed that the majority does not know the species of dog intestinal parasites, the mechanisms of transmission, the risk factors for zoonotic infections, and specific prophylactic measures. The predominance of zoonotic species in dogs in the studied region, associated with the elevated degree of misinformation of the owners, indicates that the risk of zoonotic infection by canine intestinal parasite may be high, even in one of the most developed regions of Brazil.

This paper studies a development of deterministic signal control system in urban road networks from the viewpoint of a digital control. The signal control system of the congestion length is described by a nonlinear time-varying discrete dynamic system and synthesized by using the feedback control based on the volume balance at each signalized intersection. The balance control algorithm and network control algorithm in which the three signal control parameters consisting of the cycle length, green split and offset are searched systematically so as to minimize the sum of congestion lengths at signalized intersections are presented. The configuration and development procedure of the deterministic signal control system are presented based on our studies. From the comparison of congestion lengths and signal control parameters between measurement values and simulation values, it is confirmed that the signal control systems and the signal control algorithms work effectively to minimize the congestion lengths at signalized intersections.

The traffic flow dynamics is described by a nonlinear discrete dynamic system at signalized intersections based on the volume balance. The mean link travel time are analyzed according to the classification of the traffic flow, offset control and moving direction at downstream signalized intersections in urban road networks. The mean link travel time are measured by the average rate of speed of the traffic stream which is one of test-car techniques. The mean link and OD travel time are evaluated for four routes in Fukuyama city, Japan according to their analyses. A dynamic route search algorithm which plays an essential role in the dynamic route guidance system is presented using the Dijkstra's algorithm.

A 74-year-old man who had previously undergone prosthetic graft replacement of the total aortic arch using the elephant trunk technique and of the abdominal aorta was admitted to our hospital for surgical treatment of descending aortic aneurysm. Computed tomography (CT) on admission revealed descending aortic aneurysm of 6.5 cm in diameter, and the previously placed prosthetic graft was detected in the aneurysm. Surgery for the descending aorta was performed under femoro-femoral partial bypass. During the operation, a balloon occlusion catheter introduced through the right brachial artery into the 'elephant trunk' graft was inflated before the aneurysm was opened, then the previously placed prosthetic graft was cross-clamped and the descending aorta was replaced with a new prosthetic graft with usual fashion. The postoperative course was uneventful.

It has been slightly over ten years since the development of a way to produce macroscopic quantities of fullerene, and the related discovery of fullerene nanotubes. As a result, over 1500 worldwide patents have been filed for the production and applications of these new materials. These applications are so wide ranging that they extend across different industries with products from additives to polymers, photoconductors, photo-resists, and bio-active agents to cosmetics. MER Corporation in Tucson, Arizona joined the ranks of fullerene enthusiasts at the beginning of its discovery by immediately licensing the Huffman-Kratschmer patents. While we are widely recognized as a producer of fullerene and nanotubes, MER has also been active in developing applications for fullerenes and nanotubes. The different applications investigated by MER will be reviewed in subsequent chapters. The overriding factor for the success of any of these applications, however, is the price of fullerenes. However, the price cannot come down markedly until large-scale applications are found. To introduce the first large-scale application an organization had to take a leap of faith and initiate the large-scale low-cost production. Mitsubishi/FIC Corporations has been a leader and pioneer in recognizing the need to support large-scale production effort to realize the fullerene and nano-technology commercialization dream. It is now our opportunity to realize the commercial applications. The present status of the scale-up production effort of fullerenes and the different nanotubes will be presented in this chapter.

Novel and high-performance anode materials based on modified fullerene materials, for lithium-ion rechargeable batteries were developed. The technical feasibility of lithium-ion intercalation and de-intercalation based on the degree of anion or atomic modification of the fullerene was examined. In this approach the fullerene molecule is viewed as a large anchor molecule to which various anion or atoms can be attached to form C 60 A x , where A could be hydrogen. The lithium-ion intercalation and de-intercalation could be driven by the formation of C 60 AxLix. This could result in anode materials with a potential capacity of >1200 mAh/g. Fullerenes (mixed C 60 /C 70 , pure C 60 , and pure C 70 ) and hydrogenated fullerenes, both as thin film and as pasted powdered electrode in solid polymer and liquid electrolyte, were investigated. The results of this investigation demonstrated that (1) capacities of a thin film fullerene electrode in a solid polymer electrolyte were found to be low, corresponding to three lithium ions per fullerene molecule, (2) thin film hydrogenated fullerene electrodes resulted in a significant increase in capacity, and lithium-ion intercalation was found to correspond exactly to the degree of hydrogenation of fullerenes, and (3) only certain hydrogenated fullerenes were, surprisingly, found to be effective as high-performance anode materials in liquid electrolyte. Capacities as high as 1100 mAh/g of hydrogenated fullerenes were achieved. This capacity was again found to correspond to the degree of fullerenes hydrogenation of this specific fullerene. In addition, it was found that hydrogenated fullerenes can be added in small quantities to commercial carbon to significantly reduce the irreversible capacity and improve their reversible capacity.

This paper addresses the fuzzy control problem for a class of discrete-time nonlinear systems with disturbance. In case not all the system states are available, state observer is proposed. Consequently, the whole system behavior can be attributed to a kind of the standard singularly perturbed form. The system design is based on the estimated values from the state observer. It is shown that the proposed fuzzy controller guarantees the output of the considered system follows the desired value while maintaining all signals involved in the system stable.

Finding salient features is one of the most important steps in designing a pattern recognizer. Although this issue has long been studied from the early days of research on pattern recognition, no study has yet provided a well-formalized satisfying solution which is able to be easily used in practical conditions. In this paper, we discuss in detail a new family of discriminative feature design methods, namely, discriminative feature extraction, the minimum error learning subspace method and discriminative metric design. Tracing the development of these methods clarifies a desirable research direction toward a solution to the problem.

This study examined the relationship among growth, steroid production and transforming growth factor-beta 1 (TGF-beta 1) immunolocalization in the mouse follicles cultured in vitro to evaluate the hypothesis that normally developing follicles should express TGF-beta 1 in the granulosa cells around the time of antrum formation. Preantral follicles with 151-175 microns (large category) and 125-150 microns (small category) of initial diameters were used as models for normal and retarded follicles, respectively. Growth rate and timing of antrum formation in both categories were comparable to those of in-vivo grown follicles. At the time of antrum formation, follicular diameters were similar between the two follicle categories; however, antral follicles from the large category showed larger number of granulosa cells, higher estradiol production and proportion of follicles with TGF-beta 1 positive granulosa cells. Two days after antrum formation, there were no differences in the number of granulosa cells and the proportions of follicles with TGF-beta 1 positive granulosa or theca cells between the two categories. Temporal association in large follicles between the increase in estradiol production and proportion of follicles with TGF-beta 1 positive granulosa cells at the time of antrum formation supports our hypothesis. Furthermore, this study demonstrated the usefulness of the follicle culture system in the investigations of follicular physiology.

Ovulated mouse oocytes denuded of their cumulus cells, were vitrified in a solution containing 7 M ethylene glycol as the sole cryoprotectant using one or two steps of exposure before vitrification and were diluted in 1 M sucrose solution in 5 or 10 min after warming. The results proved that the viability of oocytes are detrimentally affected by exposure to the vitrification solution even without vitrification. At 5 min dilution time, the two-step exposure was superior to the one-step in terms of the post-warming recovery rate of vitrified oocytes with normal morphology and their subsequent development to the blastocyst stage (p0.01) after fertilization in vitro. At 10 min dilution time, no significant difference between one- or two-step exposure was found. The effect of the addition of 0.5 M sucrose to the vitrification solution was also determined and did not result in a significant improvement in the viability of oocytes vitrified in one-step and diluted for 10 min. In conclusion, the results in this study indicate that oocytes can be vitrified with 7 M ethylene glycol as the sole cryoprotectant in the vitrification solution, and that the recovery of normal oocytes after one-step exposure in the vitrification solution can be improved by 10 min dilution time. However, the improvement in the recovery rate of oocytes with normal morphology and their subsequent developmental in vitro was not improved by the addition of 0.5 M sucrose to the vitrification solution.

Mouse metaphase II (M II) oocytes were exposed to 50 micrograms/microliters etoposide (ETO) before and after parthenogenetic activation with 7% ethanol and they were washed with 0.75 M sucrose. The ETO treated parthenogenetically activated oocytes were cultured or fused to single blastomeres of late 2-cell stage mouse embryo to test their ability to support development in vitro. In parallel untreated parthenogenetically activated oocytes were cultured to serve as control. None of ETO treated oocytes developed beyond the 2-cell stage, whereas 4% of the reconstituted embryos and 35% of control developed to blastocysts. It is concluded that mouse M II oocytes can be functionally enucleated by ETO treatment and can be used for nuclear transfer experiments.

To develop a method for the cytogenetic evaluation of preimplantation embryos using nonradioactive centromeric probes for chromosomes 1, 16 and X.The embryos used for this study were either fragmented or polyploid embryos rejected from an in vitro fertilization program. Prior to in situ hybridization, the embryos were treated with 0.5% protease. After application of gradual fixation, conventional hybridization protocol was followed.Ten of 11 embryos showed hybridization signals suggesting that the success rate of in situ hybridization of human embryos is improved when a modified method of digesting the zona pellucida and gradual fixation with removal of the cytoplasm are used.The method described in this study demonstrates that the zona pellucida is the key to successful in situ hybridization of whole human embryos. When the zona pellucida is removed, penetration by a probe becomes possible.

To investigate whether reflex tearing measured by the Schirmer test with nasal stimulation is better correlated with lymphocytic infiltration of the lacrimal gland and is thus a more appropriate test for Sjögren's syndrome (SS).272 patients with dry eye (age 55.5 +/- 13.5 years) with regular Schirmer test results10 mm were recruited for the study. They were divided into 2 groups depending on the results of the Schirmer test with nasal stimulation. 24 age and sex matched patients from each group were selected for lacrimal gland biopsy and 10 patients for salivary gland biopsy, and the levels of systemic autoantibodies and degrees of ocular surface staining and lymphocytic infiltration of the lacrimal and salivary glands were compared.Of the 272 patients, 187 (68.8%) had good reflex tearing (GT) and 85 (31.2%) had poor reflex tearing (PT). There were no significant differences in the age or sex of the 2 groups. Lymphocytic infiltration of the lacrimal gland was observed in 7 GT cases and 22 PT cases out of 24 cases. Lymphocytic infiltration of the salivary gland was also higher in the PT (6 of 10) than the GT group (2 of 10).Evaluation of reflex tearing by the Schirmer test with nasal stimulation identifies 2 groups of patients with dry eye. Those with poor reflex tearing were more likely to have autoantibodies and lymphocytic infiltration of the exocrine glands consistent with SS.

To determine the diagnostic value in Sjögren's syndrome (SS) of evaluating the presence of focal adenitis in labial salivary gland (LSG) and lacrimal gland (LG) biopsy specimens.We studied 105 cases with suspected SS and performed biopsies prospectively. The diagnostic criteria of Fox, et al were followed. The Chisholm-Mason grading standard and Greenspan focus score were used for evaluation.Epimyoepithelial islands and severe lymphocyte infiltration with germinal centers were observed only in LG biopsy specimens (5.9 and 3.5%, respectively). The lymphocytic focus scores of LG (2.5 +/- 2.1/4 mm2) were significantly higher than those of LSG specimens (1.3 +/- 1.1/4 mm2) in 23 cases with both biopsy results (p0.05). Six of the 32 LSG biopsies were normal while their LG biopsies were significantly higher. Conversely, 5 of the 32 LG biopsies were aided by corresponding LSG biopsies, since in LG biopsies no glandular tissue was obtained. Evaluation of both specimens for the same patient was significantly more effective in diagnosing SS than using either specimen alone (p0.01).LG biopsy specimens have a more evident histopathology than LSG biopsy specimens. We recommend that both LSG and LG biopsies be performed in patients with suspected SS to reduce false negative results and improve diagnostic accuracy.

The expression pattern of beta 1 integrin on the surface of tumor cells in patients with multiple myeloma (MM) is reviewed and compared with that of other B cell malignancies. The expression pattern of beta 1 integrin of plasma cells of healthy individuals was also compared with that of malignancies of plasma cells. Normal immature CD10+ B cell precursors in bone marrow are alpha 4+ and alpha 5+, while mature peripheral B cells are alpha 4+ and alpha 5+. In contrast to mature peripheral B cells, it is reported that plasma cells are alpha 4+ and alpha 5+. There are three points following in the phenotype characteristic of MM cells; (i) MM cells are alpha 4 strong positive, (ii) MM cells are beta 1 strong positive and (iii) MM cells are alpha 6strong positive. Because alpha 6+ cell lines of malignant plasma cells showed spread and chemotaxis on stimulation with laminin, alpha 6 beta 1 integrin might contribute of MM cells to transit laminin rich basement membrane of blood vessel to exit to the extravascular space. The difference of the phenotype between plasma cells and MM cells is the expression of alpha 5; plasma cells express alpha 5, but MM cells are lost in one half of the cases. The functional role of VLA-5 in MM cells is reviewed.

We have examined 80 patients with systemic lupus erythematosus. Of 127 head MRI studies 75 showed parenchymal changes. Brain MRI findings were classified roughly into six groups: multiple high-intensity spots on T2-weighted images, 28 studies; infarction, 33 studies; transient high-intensity lesions on T2-weighted images, 8 studies; sinus thrombosis, 3 studies (1 patient); diffuse intracranial calcification, 2 studies; inflammation, 1 study. Significant differences were noted between MRI positive and MRI negative patients with regard to age and duration of the disease. Patients with MRI lesions were significantly older than MRI negative patients, and had a longer duration of the disease. There was no significant difference in the two groups with regard to lupus anticoagulant and the level of anticardiolipin antibody.

We investigated the relationship between the amounts of Cry j I in house dust and airborne Cryptomeria japonica pollen in the same location. Cry j I was still detected in house dust collected two weeks after airborne C. japonica pollen had disappeared. Disappearance of Cry j I in house dust coincided with the disappearance of symptoms in the C. japonica pollinosis patients who lived in the same area. Airborne Cupressaceae pollen appeared during the latter half of the C. japonica pollen season. Disappearance of Cupressaceae pollen did not coincided with the disappearance of the symptoms in C. japonica pollinosis patients. Therefore, some symptoms of C. japonica pollinosis patients after C. japonica pollen disappeared from the air may be caused by pollen which had attached to clothes and been brought indoors.

To clarify the effect of respiratory muscle fatigue on ventilatory response to carbon dioxide, we performed CO2 rebreathing study before and after diaphragmatic fatigue in nine healthy males. Diaphragmatic fatigue was induced by inspiratory resistor loading and confirmed by the increase in Tension Time Index and the decrease in Pdi max at FRC. The effects of diaphragmatic fatigue were as follows: 1) S and B value of VE-CO2 curve did not change. 2) P1-CO2 curve shifted to the left but the slope of the curve did not change. 3) delta Ppl response to CO2 decreased, but delta Pdi response to CO2 did not change. 4) The increase in respiratory accessory muscle EMG was more prominent, compared to diaphragmatic EMG. 5) Rib cage movement became more marked. In conclusion, diaphragmatic fatigue (with 60 percent decrease in Pdi max at FRC) does not affect on ventilatory response to carbon dioxide. To maintain the homeostasis of the chemical ventilatory feedback system, diaphragmatic dysfunction is compensated by the increased activity of respiratory accessory muscles with possible increase in neural drive.

Airborne pollens collected in a pollen collector (Virtual Impactor) was treated with a fluorescein isothiocyanate-labeled monoclonal antibody (KW-S10) which was strictly specific to Japanese cedar pollen antigen (Cry j I). Flow cytometric analysis revealed that the intensity of fluorescence of the pollen samples treated with the antibody was greater than that of non-treated reference pollen or the antibody treated Hinoki-cypress pollen. By use of this method, it may be possible to display the airborne pollen concentration within 20 min after sampling.

We have developed an enzyme-linked immunosorbent assay (ELISA) using two anti-Cry j I monoclonal antibodies (KW-S10 and KW-S131) for the quantitation of the major allergen from Japanese cedar (Cryptomeria Japonica) pollen, Cry j I. Polystyrene microplates coated with KW-S131 were incubated with pollen allergen extracts. Cry j I, which bound to the antibody, was detected with alkaline phosphatase-conjugated KW-S10 using chromogenic enzyme substrate. Cry j I could be measured in concentration of between 0.16 and 2.5 ng/ml by this assay. Intra- and inter-assay coefficients of variation were 1.1-3.5% and 0.9-4.6%, respectively. This assay was considered that it was specific for Cry j I of Japanese cedar pollen because it didn't react with allergens of hinoki (Chamaecypairs obtusa) pollen which have antigenicity in common to Japanese cedar pollen. This assay would be useful for the standardization of Japanese cedar pollen allergen extracts.

We experienced malignant localized mesothelioma of which origin was visceral pleura. According to, 1) the preoperative chest X-ray and chest CT which showed extra-pleural sign, 2) the rapid tumor growth, and 3) the result from needle biopsy, we diagnosed malignant localized mesothelioma of which origin was parietal pleura. Surgical treatment was performed, and diagnosed that its origin was visceral pleura. The tumor invaded the lung. It is dangerous to diagnose by means of needle biopsy because of malignant cell implantation. We recommend that firstly the surgical treatment should be carried out for malignant mesothelioma, which needs extended resection for preventing its recurrence.

We produced monoclonal antibodies against the major allergen of Japanese cedar (Cryptomeria japonica) pollen, Cry j I. KW-S10 antibody reacted only with Japanese cedar pollen and KW-S91 antibody reacted to most angiospermae pollens as well as Japanese cedar pollen. Using these antibodies, we devised a new counting method of Japanese cedar pollen allergen particles by an immunoblotting technique. Airborne pollen allergens were collected on vaseline coated glass slides or Burkard's sampling tape and were transferred onto polyvinylidene difluoride (PVDF) membranes. The membranes were treated with anti-Cry j I monoclonal antibody conjugated with alkaline phosphatase. Pollen allergens were detected as spots on the membranes after staining with phosphatase substrate (BCIP/NBT). This method using KW-S10 antibody measured only the amounts of allergen from Japanese cedar pollen, while with KW-S91 antibody, the method measured the amounts of pollens which have antigenicity in common with Japanese cedar pollen.

This study examined the effect of experimental diabetes mellitus on the dorsal tongue of rats which were made diabetic by tail-vein injections of streptozotocin (50 mg/kg) and then raised for either 5 or 10 months. Lingual papillae were observed by scanning electron microscopy (SEM) and light microscopy. Morphological changes in lingual mucosal capillaries in 10-month diabetic rats were observed by electron microscopy (TEM). In the study of cellular movement in the lingual dorsal epithelium, bromodeoxyuridine (BrdU) was applied as a tracer for studying DNA replication. In diabetic rats, lingual papillae showed morphological atrophic changes. The lingual mucosal capillaries' alterations included endothelial cells with numerous cell projections into the lumen, degenerated cell organs, increased basement membrane width, and narrowed capillary lumen. BrdU labeling index among the basal cells was reduced in diabetic rats which indicates a possible retardation of their epithelial-tissue activity. In diabetes mellitus, direct metabolic disturbances to the epithelia because of insulin deficiency first occurred, successively diabetic microangiopathy appeared on the lingual mucosal capillaries. The appearance of diabetic microangiopathy caused tissue hypoxia, which induced atrophic changes to the epithelia.