The relationship between lip-closing force and dental arch morphology in patient with Angle Class I malocclusion.

Dental arch morphology and tooth position are affected by lip-closing force (LCF). This study aimed to quantitatively evaluate the relationships between the horizontal or vertical balance of the LCF generated during maximum voluntary pursing-like movements and dental arch length (DAL) or width (DAW) or the lingual inclination of the upper or lower 1st molars (LIUM, LILM) in patients with Angle Class I malocclusion. Sixteen subjects with Angle Class I malocclusion (median age: 23·4 ± 5·9 years) who had never undergone orthodontic treatment were randomly selected. LCF was measured in eight directions during maximum voluntary pursing-like lip-closing movements. Dental arch models were scanned and analysed to obtain DAW, DAL, LIUM and LILM measurements. Mandibular deviation was measured on posteroanterior cephalograms. A significant negative correlation was detected between maxillary DAL and upper LCF. Maxillary DAL, DAW and the DAL/DAW ratio displayed significant negative correlations with total LCF and upper LCF. However, no significant correlations were detected between any mandibular dental arch morphological parameter and LCF. The difference in the LIUM between the deviation and non-deviation sides exhibited a significant positive correlation with the difference in upper LCF between the deviation and non-deviation sides and was significantly negatively correlated with the difference in lower LCF between the deviation and non-deviation sides. These results suggest that upper LCF is related to maxillary DAL, and the horizontal balance of the LCF of the upper and lower lips is related to the LIUM during pursing-like lip-closing movements in patients with Angle Class I malocclusion.

Note: Application of CR-39 plastic nuclear track detectors for quality assurance of mixed oxide fuel pellets.

A CR-39 plastic nuclear track detector was used for quality assurance of mixed oxide fuel pellets for next-generation nuclear power plants. Plutonium (Pu) spot sizes and concentrations in the pellets are significant parameters for safe use in the plants. We developed an automatic Pu detection system based on dense α-radiation tracks in the CR-39 detectors. This system would greatly improve image processing time and measurement accuracy, and will be a powerful tool for rapid pellet quality assurance screening.

Operational test of micro-oven for 48Ca beam.

In order to supply a high-intensity and stable (48)Ca beam from the RIKEN 18-GHz electron cyclotron resonance ion source, we are conducting operational tests of a micro-oven. A mixture of CaO and Al powders is placed into the crucible of the micro-oven and heated to produce metallic calcium by a reductive reaction. The successful production of a calcium beam was confirmed. In addition, we reduced the material consumption rate by using a so-called "hot liner," and we enhanced the beam intensity by applying a negative voltage bias to the micro-oven, the effect of which is similar to the effect of a "biased disk."

Genetic analysis of novel avian A(H7N9) influenza viruses isolated from patients in China, February to April 2013.

Novel influenza viruses of the H7N9 subtype have infected 33 and killed nine people in China as of 10 April 2013. Their haemagglutinin (HA) and neuraminidase genes probably originated from Eurasian avian influenza viruses; the remaining genes are closely related to avian H9N2 influenza viruses. Several characteristic amino acid changes in HA and the PB2 RNA polymerase subunit probably facilitate binding to human-type receptors and efficient replication in mammals, respectively, highlighting the pandemic potential of the novel viruses.

Establishment of an ELISA system for the determination of Japanese monkey calreticulin and its application to plasma samples in macaques.

BACKGROUND: Calreticulin (Crt) is a molecular chaperone in endoplasmic reticulum, assisting a correct folding of glycoproteins. Establishment of its assay method might be advantageous to determine the Crt level in cell or other biosystems. METHODS: An enzyme-linked immunosorbent assay (ELISA) system for the determination of Crt of Japanese monkey, Macaca fuscata, was developed in this study. Japanese monkey Crt protein expressed in Escherichia coli was used as a standard protein. RESULTS AND DISCUSSION: The assay was sensitive even to <10 ng/ml of Crt. Since the amino acid sequence of Crt is quite similar (99%, similarity) between the Japanese and rhesus monkeys, the ELISA was applied to the determination of plasma Crt in these two species in association with various diseases. The Crt level increased significantly in monkeys suffering from pneumonia and diarrhea, suggesting that the ELISA might be applicable for preliminary diagnosis of inflammatory disease.

Development-dependent expression of calreticulin in the brain and other tissues of the Japanese monkey, Macaca fuscata.

BACKGROUND: Calreticulin (CRT) is a molecular chaperone localized to endoplasmic reticulum lumen. The protein is involved in the correct folding of glycoproteins and its cellular level changes depending on various physiologic conditions. METHODS: To clarify the Crt level in various tissues of the Japanese monkey during postnatal development, an enzyme-linked immunosorbent assay system that we have established was applied. RESULTS AND CONCLUSIONS: Calreticulin is distributed ubiquitously in various tissues. Its level in the heart, lung, liver, spleen, and kidney was high in newborns and decreased in juveniles and adults. In various cerebral areas, Crt was present in the gray matter but scarcely found in the white matter. The Crt levels in the cerebral areas were low in newborns and increased in juveniles and adults. These distribution and developmental changes in Crt might be correlated with the quality control of glycoproteins that are synthesized in respective tissues.

Molecular cloning and gene expression of endoplasmic reticulum stress proteins in Japanese monkey, Macaca fuscata.

BACKGROUND: Immunoglobulin heavy-chain binding protein (BiP), calreticulin (Crt), and protein disulfide isomerase (PDI), are major resident endoplasmic reticulum (ER) stress proteins which are involved in diverse roles relating to successful folding, assembly, intracellular localization, and degradation of other proteins. METHODS: In this study, we molecular cloned cDNAs for BiP, Crt, and PDI from Japanese monkey (Macaca fuscata), and analyzed tissue-specific expression of respective genes. RESULTS AND CONCLUSIONS: The lengths of protein-coding regions of these cDNAs for BiP, Crt and PDI are 1965, 1254, and 1533 bp, respectively. Each protein has a signal peptide and a KDEL motif in N- and C-terminal parts respectively, showing its intracellular localization to be the lumen of the ER. These stress proteins are highly conserved, showing that their similarities among mammals are more than 90% in the level of amino acid. The expression of the genes for stress proteins differed among the monkey tissues examined. BiP and PDI gene expression was predominant in secretory tissues such as liver and kidney, and brain tissues. But Crt gene expressed rather ubiquitously in a variety of tissues.

Effect of vascularity on canine distracted tibial callus consolidation.

In the consolidation period of distraction osteogenesis, mineralization occurs before corticalization. We hypothesized that the increased rate of bone mineral density correlates to the density of vascularity in the callus. We unilaterally lengthened the tibia in eight adult beagles. After a waiting period of 7 days, tibiae were lengthened for 30 days. After a consolidation period of an additional 60 days, all animals were euthanized. Just before euthanasia, blood vessels were perfused with 50% w/v barium sulfate solution, and soft radiographs of the distracted callus and the control tibiae were taken. Bone mineral density of the regenerated bones was measured preoperatively by quantitative computed tomography on Days 37, 68, and 98. Increases of the percent bone mineral density (from Day 37 to Day 98 and from Day 68 to Day 98) correlated with the blood vessel volume density ratios on Day 98. Our results suggest that preservation of the higher density of blood vessels in the consolidation period could lead to the better mineralization of the distracted callus.

Cleavage activity of the sapovirus 3C-like protease in Escherichia coli.

We recently determined the ORF1 cleavage map of Mc10, a human sapovirus (SaV) strain, as follows: NH2-p11-p28-p35(NTPase)-p32-p14(VPg)-p70(Pro-Pol)-p60(VP1)-COOH. This cleavage was dependent on the viral encoded 3C-like protease. To identify the cleavage site of SaV ORF1, putative p70 (Pro-Pol) and p14-p70 (VPg-Pro-Pol) were expressed as N-terminal GST and C-terminal 6 x His-tag fusion proteins in Escherichia coli, and the expressed products were analyzed by SDS-PAGE and Western blotting. Our results indicated that the efficient proteolytic cleavage occurred between p14 (VPg) and p70 (Pro-Pol), and N-terminal amino acid sequencing revealed that the cleavage site was between E(1055) and A(1056). In contrast, the p70 (Pro-Pol) was not further cleaved. We also found that SaV protease cleaved the Q/G site within the rhinovirus 3C protease recognition site. Site-directed mutagenesis in a conserved GDCG motif of the protease completely abolished these proteolytic activities. This is the first report to identify the cleavage site of the SaV ORF1 polyprotein.

Visual results and complications of temporal incision phacoemulsification performed with the non-dominant left hand by junior ophthalmologists.

AIMS: To assess the results of temporal incision phacoemulsification and aspiration performed with dominant and non-dominant hand of ophthalmology trainees. METHODS: Retrospective analysis were made of 203 surgeries with dominant hand and 207 with non-dominant by five trainees at two institutions. Trainees sat at the patient's head, manipulating instruments with the dominant right hand for the right eye, and the non-dominant left hand for the left eye. RESULTS: Vitreous loss occurred in 12 (5.9%) of 203 dominant operated eyes and seven (3.4%) of 207 non-dominant operated eyes. The rate of endothelial cell loss was 6.1% (9.8%) in dominant and 7.4% (12.4%) in non-dominant. Mean ultrasound time were 1.81 (0.70) minutes in dominant and 1.78 (0.78) minutes in non-dominant. One trainee showed statistically significant excesses in incidence of vitreous loss in dominant operated eyes (8.7%, p=0.0270), and one showed statistically significant prolongation of the operation in nondominant operated eyes (26.3 minutes, p=0.0315). In all other trainees, all parameters had no difference in both sides. CONCLUSIONS: Ophthalmology trainees could successfully learn the technique with both hands. The authors consider that the skill of the non-dominant hand may be knowledge based and that surgeons avoid mistakes by mental efforts.

Yersinia pseudotuberculosis infection in breeding monkeys: detection and analysis of strain diversity by PCR.

In the last three decades, several monkeys reared in outdoor/indoor-outdoor breeding colonies and cages of the Primate Research Institute, Kyoto University, died of yersiniosis caused by Yersinia pseudotuberculosis, necessitating introduction of a method to detect the bacteria rapidly and thus allow preventive measures to be undertaken. A rapid nested polymerase chain reaction (PCR) method for identification of Y. pseudotuberculosis in fecal samples and a random amplified polymorphic DNA (RAPD)-PCR approach for distinguishing between bacterial strains were therefore developed. Yersinia pseudotuberculosis isolates from monkey specimens were found to be classifiable into several types. To determine the source of infection, hundreds of fecal samples of wild rats, pigeons, and sparrows were collected from around the breeding colonies and cages, and subjected to PCR analyses. Yersinia pseudotuberculosis was detected in 1.7% of the fecal samples of wild rats. The DNA fingerprints of the bacteria revealed by RAPD-PCR were the same as that of one strain isolated from macaques, suggesting the wild rat to be a possible source of infection.

Pepsinogens, progastricsins, and prochymosins: structure, function, evolution, and development.

Five types of zymogens of pepsins, gastric digestive proteinases, are known: pepsinogens A, B, and F, progastricsin, and prochymosin. The amino acid and/or nucleotide sequences of more than 50 pepsinogens other than pepsinogen B have been determined to date. Phylogenetic analyses based on these sequences indicate that progastricsin diverged first followed by prochymosin, and that pepsinogens A and F are most closely related. Tertiary structures, clarified by X-ray crystallography, are commonly bilobal with a large active-site cleft between the lobes. Two aspartates in the center of the cleft, Asp32 and Asp215, function as catalytic residues, and thus pepsinogens are classified as aspartic proteinases. Conversion of pepsinogens to pepsins proceeds autocatalytically at acidic pH by two different pathways, a one-step pathway to release the intact activation segment directly, and a stepwise pathway through a pseudo-pepsin(s). The active-site cleft is large enough to accommodate at least seven residues of a substrate, thus forming S4 through S'3 subsites. Hydrophobic and aromatic amino acids are preferred at the P1 and P'1 positions. Interactions at additional subsites are important in some cases, for example with cleavage of kappa-casein by chymosin. Two potent naturally occurring inhibitors are known: pepstatin, a pentapeptide from Streptomyces, and a unique proteinous inhibitor from Ascaris. Pepsinogen genes comprise nine exons and may be multiple, especially for pepsinogen A. The latter and progastricsin predominate in adult animals, while pepsinogen F and prochymosin are the main forms in the fetus/infant. The switching of gene expression from fetal/infant to adult-type pepsinogens during postnatal development is noteworthy, being regulated by several factors, including steroid hormones.

Novel mutations in the a3 subunit of vacuolar H(+)-adenosine triphosphatase in a Japanese patient with infantile malignant osteopetrosis.

A case of infantile malignant osteopetrosis is described. The patient died from respiratory hemorrhage at 7 months of age despite treatment that included high doses of active vitamin D and administration of interferon-gamma. A postmortem examination revealed the presence of many osteoclasts in the bone, which lacked ruffled borders. This observation was consistent with the histology of bone reported in Atp6i-knockout mice, which lack the gene encoding the a3 subunit of vacuolar-type H(+)-adenosine triphosphatase (ATPase). Sequence analysis of the TCIRG1 gene encoding the a3 subunit revealed two novel mutations: a deletion/insertion mutation in exon 9 and a T-to-C transition at the splice donor site of intron 19. The former mutation caused a frame shift and premature stop codon. The latter was associated with abnormal splicing, which was confirmed by sequencing the products amplified by reverse transcription-polymerase chain reaction (RT-PCR), using total RNA from the liver specimen as template. Although several mutations in the TCIRG1 gene in infantile malignant osteopetrosis have been reported in other populations, this is the first case of a Japanese patient with a mutation identified in this gene. These results support the important role of the subunit in the function of the proton pump.

Phylogenetic position of Eulipotyphla inferred from the cDNA sequences of pepsinogens A and C.

Although to date the phylogenetic position of the provisional order Eulipotyphla has been assessed by various molecular markers, it has not been conclusively clarified due to low statistical supporting values and inconsistent results. To clarify the phylogenetic position of Eulipotyphla, we cloned cDNAs for pepsinogens A and C from five mammalian species belonging to four different orders and determined their nucleotide sequences. Molecular phylogenetic analysis based on the 1st and 2nd codon positions of the protein-coding region of cDNA sequences strongly supported the close relationship between Eulipotyphla and Chiroptera. Carnivora was found to be a sister group to these two orders. The monophyly of the order Rodentia and that of the cohort Glires (Rodentia and Lagomorpha) was also shown by the present phylogenetic trees of pepsinogens.

Results of vitrectomy performed at the time of phacoemulsification complicated by intravitreal lens fragments.

AIM: To evaluate outcome of vitrectomy performed at the time of phacoemulsification complicated by intravitreal lens material. METHODS: Clinical records associated with consecutive 8536 phacoemulsification procedures were reviewed retrospectively. RESULTS: 17 (0.20%) eyes had a posterior capsule rupture with retained lens material in the vitreous cavity that required vitrectomy. Final visual acuity was 0.5 or better in 14 eyes (82%) and 0.4 to 0.1 in three eyes (18%). Retinal detachment occurred in one eye during vitrectomy and two after the surgery. Cystoid macular oedema was observed in two eyes and none developed glaucoma. The corneal endothelial cell loss was 5.7% (SD 6.8 %) (n=15) at 3-6 months postoperatively. CONCLUSIONS: Combined vitrectomy and intraocular lens implantation at the time of phacoemulsification complicated by intravitreal lens material is an option to be considered to reduce the risk of postoperative complications including secondary glaucoma and corneal endothelial cell damage.

Molecular characterization of hepatitis C virus genotype 2a from the entire sequences of four isolates.

Genotype 2a hepatitis C virus (HCV) has different characteristics from genotype 1b, such as responsiveness to interferon therapy. Such type-specific characteristics appear to be due to differences in the HCV genome sequence. The complete sequences of genotype 2a HCV genome isolated from four patients with chronic hepatitis C were determined, and nucleotide and deduced amino acid sequences were compared within genotype 2a, as well as between genotype 2a and 1b. Whereas the amino acid sequence similarity of the core region was highest within genotype 1b, the NS3 and NS4B regions of exhibited greater similarity than the core region in genotype 2a. The serine protease and helicase motifs in the NS3 region were well conserved in genotype 2a to the same degree as in genotype 1b. However, the putative secondary structure of 2a isolates was significantly different from that of the 1b isolates. Analysis of amino acid similarity between genotypes 2a and 1b revealed the lowest degree of similarity in the E1 region, followed by the NS2 and NS5A region. Sequences of genotype 2a in the interferon-sensitivity determining region (ISDR) located in the NS5A region had a deletion of four amino acids compared with that of genotype 1b. When the ISDR of the genotype 2a was aligned for maximal similarity, it exhibited similarity of only 52.5-55.0% when compared with that of HCV-J, which belongs to genotype 1b. These findings for the entire sequences of genotype 2a isolates will contribute to virological studies of HCV.

[Inhibition of HCFU absorption after resection for gastric cancer--application of hydroxyaluminium gel].

HCFU (carmofur; Mifurol) is an 5-FU analog. The maximum blood concentration of HCFU in HCFU fraction (Cmax) after gastric resection was higher than before resection. Hot sensation and pollakiuria, characteristic side effects of HCFU, are dependent on concentration of HCFU fraction in blood. Therefore, it is considered that the frequency of occurrence of side effects after gastric resection is high. For that reason, we thought that if absorption of HCFU could be reduced, fewer side effects would result. We focused on the fact that drugs which include aluminium gel may decrease absorption in combined drugs, and thought it would be possible to delay absorption in HCFU by using them. We studied the HCFU concentration in the HCFU fraction and 5-FU concentration in blood, respectively, in two cases: 1) single oral administration of HCFU 100 mg and 2) coadministration with hydroxyaluminium gel (ALG) 10 ml in the whole or partial resection of gastric cancers for 8 patients. We found that the concentration of HCFU in the HCFU fraction 2 hours after its administration decreased significantly: 3.24 +/- 1.78 (single administration), 1.37 +/- 0.91 (coadministration with ALG) (p = 0.023). HCFU concentration in the HCFU fraction seemed to decrease for coadministration with ALG in the area under the time-blood concentration curve (AUG) (p = 0.071). The 5-FU concentration did not seem to decrease in either case. From these results, the coadministration of HCFU with ALG seems to be effective for the inhibition of adverse drug reaction after the resection of gastric cancers.

Estimated sleep debt and work stress in Japanese white-collar workers.

Sleep debt on weekdays in Japanese white-collar workers, estimated using a questionnaire, was associated with age, overtime, and self-rated workload.

Down-regulation of translation driven by hepatitis C virus internal ribosomal entry site by the 3' untranslated region of RNA.

The genome of hepatitis C virus (HCV) is a single-stranded RNA of positive polarity that has a poly(U/C) tract followed by a highly conserved 98-nt sequence, termed the X region, in the 3' untranslated region (UTR). To investigate the effect of the 3'UTR on the HCV translation that depends on the internal ribosomal entry site (IRES), we prepared a deletion HCV RNA, MA delta, that lacked the RNA region from nt 1286 to 8785. A series of MA delta RNAs that differ in the primary structure of their 3'UTR, were generated and examined for their translation efficiencies in reticulocyte lysates. Deletion of the poly(U/C) tract and/or stem-loop structure (SL) 3 region of 3'X resulted in enhancement of the translation efficiency. Translation of MA delta RNA was inhibited by the addition of recombinant polypyrimidine tract-binding protein (PTB). A similar inhibition by PTB, however, was observed when an RNA lacking the poly(U/C) tract or SL3 region was used. The inhibitory effect by PTB was not obvious for MA delta (1041) RNA composed of nt 1 to 1041 but MA delta (8928) RNA composed of nt 1 to 1285 and nt 8786 to 8928. These results suggest that the observed down-regulation of HCV translation by the 3'UTR is mediated by some host factor(s) other than PTB, and that a PTB site for inhibition resides in the coding sequence of nt 1042 to 8928 of MA delta RNA.