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Bilateral Sagittal split ramus osteotomy (BSSRO) represents the majority of mandibular osteotomies. Intraoperative risks associated with BSSRO include unfavorable fractures, major bleeding episodes, and inferior alveolar nerve (IAN) and lingual nerve damage. In particular, injury to the IAN during BSSRO is a well-documented complication. However, reports of immediate microneural repair of transected IAN are scarce. In this report, we describe a case of neurosensory disturbance due to transection of the IAN during BSSRO. A 20-year-old woman underwent BSSRO at our hospital. Intraoperatively, the IAN was transected during osteotomy, and immediate microneural repair of the IAN was performed combined with polyglycolic acid (PGA) sheet. We performed the stellate ganglion block (27 times), and prescribed 6.6 mg of dexamethasone sodium phosphate postoperatively from the day of the surgery for 14 days and Vitamin B12 for 6 months postoperatively. We evaluated the response to touch sensation using the Semmes-Weinstein monofilaments test (SW test) and two-point discrimination (2PD), pain sensation, and temperature sensation. The right mental nerve did not respond to any quantitative test and sensation of pain just after the operation. One year and three months postoperatively, the SW value was 2.83 and 2PD distance was 11 mm, which were similar to the values on the normal side. The patient responded to sensations of heat and cold and 2-g pain sensation similar to that before the surgery. All responses to sensations were restored to normal one year and three months postoperatively. The postsurgical evaluation revealed marked improvement and significant recovery in sensations of the IAN, following immediate neurorrhaphy combined with PGA sheet during SSRO.

Major bleeding associated with sagittal split ramus osteotomy (SSRO) involves vessels such as the inferior alveolar, facial, and maxillary arteries and veins, and the retromandibular vein (RMV). The present study aimed to clarify and classify the three-dimensional variations in RMV position and course direction in relation to the mandible. Specimens comprised a total of 15 scientific cadavers, and the relationship between RMV and the mandible lateral and posterior views was observed. We identified 3 patterns on the lateral view, the mean distance between the RMV and the posterior border of the ramus was 3.9 mm at the height of the lingula. A total of five course patterns were identified on the posterior view. In no course pattern, the RMV inferior to the lingula was lateral to its position superior to the lingual. The present findings suggest that it may be possible to predict correlations with intraoperative bleeding risk. Further study is planned using contrast computed tomography in patients with jaw deformity for skeletal classification.

Hexachlorobutadiene (HCBD) is a persistent organic pollutant (POP) that is banned in many countries. To investigate the destruction of HCBD in industrial waste, we conducted incineration tests at a commercial-scale incineration plant equipped with a 130-t/day rotary kiln and at a pilot-scale plant with approximately 2-kg/h of feeding rate. At the commercial-scale plant, waste oil containing 37% by weight of HCBD was co-incinerated at 1058 °C together with the solid and liquid industrial wastes that the plant normally processes. The concentrations of HCBD in the bottom ash (0.25 ng/g-dry), effluent slurry (2.8 ng/L), and stack gas (8.2 ng/m3N) were comparable with those obtained when only the usual industrial waste was incinerated. A destruction efficiency of 99.999996% was obtained from the co-incineration test, with negligible discharge of chlorinated dioxins and furans. At the pilot-scale plant, refuse-derived fuel spiked with HCBD was incinerated. It was found that both decomposition and unintentional generation of HCBD occurred simultaneously in the primary combustion chamber; however, the decomposition rate exceeded the generation rate to afford a destruction efficiency of 99.99993%. Together, these results indicate that HCBD contained in industrial waste can be efficiently destroyed in best available technology (BAT) rotary kiln waste incinerators.

It is mathematically proved that inference based on α-cuts and generalized mean (α-GEMII) deduces consequences converging to fuzzy sets mapped by linear fuzzy-valued functions, to be represented with α-GEMII, as the number of fuzzy rules increases. The proof indicates that α-GEMII satisfies axiomatic properties and can contribute to presenting interpretability in designing fuzzy systems in the rule base. Such properties do not hold in conventional methods based on the compositional rule of inference. Simulation results show that the difference between deduced consequences and fuzzy sets mapped by linear fuzzyvalued functions is smaller as the number of fuzzy rules increases, in which the difference is evaluated by mean square errors. The discussions may lead to improvements of the interpretability in representing nonlinear fuzzy-valued functions by using α-GEMII.

Antimicrobial peptides (AMPs) are important components of the host innate defense system against pathogenic microbial invasion in many organisms. In the present study, we cloned by RT-PCR a cDNA from total RNA prepared from the skin of the Japanese brown frog Rana japonica. The cDNA directs the synthesis of a novel, non-C-terminally alpha-amidated peptide composed of 21 amino acid residues (FLGSLIGAAIPAIKQLLGLKK). The putative peptide showed limited sequence similarity to atypical acyclic brevinin-1OK family AMPs originally isolated from the skin of the Ryukyu brown frog (R. okinavana), which lacks the COOH-terminal cyclic domain commonly observed in typical brevinin-1 groups, but that contains a C-terminally alpha-amidated residue. Although it is unclear whether the peptide, designated brevinin-1Ja, is produced in R. japonica skin, a synthetic replicate of the peptide showed differential growth-inhibiting activity against the Gram-positive bacterium Staphylococcus aureus and Gram-negative bacterium Escherichia coli (minimal inhibitory concentrations: 15 microM and 119 microM, respectively), and produced cell death of mammalian COS7 cells (LD50=28 microM).

RIKEN Nishina Center for Accelerator Based Science, Saitama 350-0198, Japan 1 Faculty of Science, Niigata University, Niigata 950-2181, Japan 2 Graduate School of Science and Technology, Niigata University, Niigata 950-2181, Japan 3 Faculty of Science and Technology, Tokyo University of Science, Chiba 278-8510, Japan 4 Graduate School of Science and Engineering, Yamagata University, Yamagata 990-8560, Japan 5 Department of Physics, Saitama University, Saitama 338-8570, Japan E-mail: daiya@riken.jp

Using a combination of reverse-transcription polymerase chain reaction and the 5'- and/or 3'-rapid amplification of cDNA ends, we cloned, from a Japanese brown frog (Rana japonica) skin total RNA preparation, cDNAs encoding biosynthetic precursors for the antimicrobial peptides (AMPs) japonicin-1Ja (FFPIGVFCKIFKTC), japonicin-2Ja (FGLPMLSILPKALCILLKRKC), and temporin-1Ja (ILPLVGNLLNDLL.NH2). These peptides were previously isolated from an extract of R. japonica skin. The present study is the first report to describe the molecular cloning of the cDNA encoding a japonicin-2 family peptide. The nucleotide and deduced amino acid sequence analyses revealed that the hypothetical precursor protein of japonicin-2Ja, as well as japonicin-1Ja and temporin-1Ja, is organized similarly to those of typical amphibian AMP precursors, with a highly conserved signal peptide, a relatively well conserved intervening sequence, and a hypervariable AMP mature region. Antimicrobial assays for synthetic replicates of cyclic and linear japonicin-2Ja revealed that the intramolecular disulfide bond is necessary for activity. A semi-quantitative analysis by real-time RTPCR using TaqMan probes revealed that the relative values of preprojaponicin-2Ja mRNA expression levels in the skin, skeletal muscle of hind leg, kidney, testis, small intestine, and stomach total RNA sample specimens in adult R. japonica were 6.5×10(5), 9.6, 2.0, 1.6, 1.6, and 1.0, respectively. The presence of preprojaponicin-2Ja mRNAs in the cytoplasm of glandular cells in R. japonica dorsal skin glands was demonstrated by means of in situ hybridization using digoxigenin-labeled cRNA probes for the precursor.

Theoretical aspects are provided for inference based on α-cuts and generalized mean (α-GEMII). In order to clarify the basic properties of the inference, fuzzy tautological rules (FTRs) are focused on, which are composed by setting fuzzy sets in consequent parts identical to those in antecedent parts of initially given fuzzy rules. It is mathematically proved that the consequences deduced with FTRs are closer to given facts as the number of FTRs increases. The aspects provided in this paper are appropriate from axiomatic viewpoints and can contribute to interpretability in fuzzy systems constructed with α-GEMII. They are not obtained in conventional methods based on the compositional rule of inference. Simulations are performed by evaluating difference (mean square errors) between given facts and deduced consequences under the condition that convex and symmetric fuzzy sets are given as facts. Their results show that the difference becomes smaller as the number of FTRs increases. Thereby, it is confirmed that α-GEMII has an advantage in the interpretability with respect to FTRs over the conventional methods.

A governing scheme is proposed for fuzzy constraint propagation from given facts to consequences in convex forms, which is applied to the inference method based on α-cuts and generalized mean. The governing is performed by self-tuning which can reflect the distribution forms of fuzzy sets in consequent parts to the forms of deduced consequences. Thereby, the proposed scheme can solve the problems in the conventional inference based on the compositional rule of inference that deduces fuzzy sets with excessive fuzziness increase and specificity decrease. In simulations, it is confirmed that the proposed scheme can effectively perform the constraint propagation from given facts to consequences in convex forms while reflecting fuzzy-set distributions in consequent parts. It is also demonstrated that consequences are deduced without excessively large fuzziness and small specificity.

Previous studies have led to the isolation of histone H2B with antibacterial properties from an extract of the skin of the Schlegel's green tree frog Rhacophorus schlegelii and it is now demonstrated that the intact peptide is released into norepinephrine-stimulated skin secretions. In order to investigate the mechanism of action of this peptide, a maltose-binding protein (MBP)-fused histone H2B (MBP-H2B) conjugate was prepared and subjected to antimicrobial assay. The fusion protein showed bacteriostatic activity against Escherichia coli strain JCM5491 with a minimum inhibitory concentration of 11 microM. The lysate prepared from JCM5491 cells was capable of fragmenting MBP-H2B within the histone H2B region, but the lysate from the outer membrane proteinase T (OmpT) gene-deleted BL21(DE3) cells was not. FITC-labeled MBP-H2B (FITC-MBP-H2B) penetrated into the bacterial cell membrane of JCM5491 and ompT-transformed BL21(DE3) cells, but not into ompT-deleted BL21(DE3) cells. Gel retardation assay using MBP-H2B-deletion mutants indicated that MBP-H2B bound to DNA at a site within the N-terminal region of histone H2B. Consequently, it is proposed that the antimicrobial action of histone H2B involves, at least in part, penetration of an OmpT-produced N-terminal histone H2B fragment into the bacterial cell membrane with subsequent inhibition of cell functions.

Two quail lines, H and L, which were developed for high (H) and low (L) antibody production against inactivated Newcastle disease virus antigen, were used to examine differences in the organization, structure and expression of the quail Mhc class IIB genes. Four Coja class IIB genes in the H line and ten Coja class IIB genes in the L line were identified by gene amplification using standard and long-range PCRs and sequencing of the amplified products. RFLP analysis, sequencing and gene mapping revealed that the H line was fixed for a single class IIB haplotype, which we have designated CojaII-02HL- CojaII-01HL. In contrast, evidence was found for two class IIB haplotypes segregating in the L line. Some individuals were found to be homozygous for haplotype CojaII-08L- CojaII-07L and others were found to be heterozygous CojaII-08L- CojaII-07L/ CojaII-02HL- CojaII-01HL. However, expression of CojaII-02HL- CojaII-01HL was not detected in the L line. SRBC immunization induced a measurable antibody response in the serum and a line-specific class IIB gene expression in the peripheral white blood cells. CojaII-01HL was expressed at the highest level in the H line and CojaII-07L in the L line. The expression of the class IIB mRNA reached the highest level at approximately 1 week after the primary antibody response and then declined exponentially. The antibody and class IIB gene expression data obtained in response to SRBC immunization provide further evidence that quails within the L line had reduced immunocompetence compared with those in the H line.

O -Methyltransferase I, which catalyzes conversions both of demethylsterigmatocystin (DMST) to sterigmatocystin (ST) and of dihydrodemethylsterigmatocystin (DHDMST) to dihydrosterigmatocystin (DHST) during aflatoxin biosynthesis, was purified to apparent homogeneity from the cytosol fraction of the mycelia of Aspergillus parasiticus NIAH-26 through the following chromatography series: phenyl-Sepharose, DEAE-Sepharose, phenyl-Sepharose, Sephacryl S-300, and Matrex gel Green A. The apparent molecular mass was estimated at 150 kDa based on Sephacryl S-300 gel filtration chromatography, and the denaturing molecular mass was 43 kDa based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The pI of the enzyme was 4.4, and the optimal pH for activity was broad, from 6.5 to 9.0. In competition experiments using the purified enzyme, the formation of ST from DMST was suppressed when DHDMST was added to the reaction mixture and DHST was newly formed. These results indicate that DMST and DHDMST commonly serve as substrates for the enzyme. The K m of the enzyme for DMST was 0.94 μM, and that for DHDMST was 2.5 μM. Interestingly, MT-I kinetics deviated substantially from standard Michaelis-Menten kinetics, demonstrating substrate inhibition at a higher substrate concentration.

The adherent chicken cell line, MDCC-MSB1-41C, was highly transplantable and metastatic in vivo, compared with the parental non-adherent cell line MDCC-MSB1 from Marek's disease (MD) lymphoblastoid tumor. For clarification of differences in extracellular matrix proteins in MSB1-41C and MSB1 cells, examination was made of various components of extracellular matrix proteins. A detachment experiment indicated the protein(s) recognizing the Arg-Gly-Asp (RGD) sequence, the minimum structure required for recognition by the cell-surface receptors, is essential for the adherent character. Immunoblot assay using antibodies showed increased expression of fibronectin, fibronectin receptors, and vinculin on MSB1-41C cell lines. RGD-directed integrins mediate important cell-cell adhesive interaction and these interactions with extracellular matrix proteins may thus possibly be requisite for migration, proliferation and metastatic dissemination of MDCC-MSB1-41C cells. The RGD-containing peptide in the culture medium could cause detachment of cultured adhesive lymphoid leukosis LSCC-1104X5 cells from the dish too.

Micrococcus rubens, a gram-positive occus, usually forms large, cubic packets of more than 500 cells that are regularly arranged in three-dimensional cell groups. In medium with extremely low concentration of Mg2+ and phosphate, in which the cells can only grow on a agar surface, it formed small groups of 2 to 20 cells. Irregularly arraged cell groups of intermediated size were obtained in culture media containing intermediated concentrations of Mg2+ and phosphate. Mutants that formed irregular cell groups of intermediate size under normal culture conditions were also obtained.

The H strain of avian erythroblastosis virus (AEV-H) was recently isolated from the liver emulsion of a chicken that suffered from erythroblastosis after being inoculated with subgroup A leukosis virus. AEV-H induced erythroblastosis or sarcoma when inoculated into chickens and transformed chick embryo fibroblasts (CEF) in vitro. Analysis of viral proteins synthesized in cells, which were named HNP, transformed by AEV-H but not producing transforming virus revealed tha the genome of AEV-H directed the synthesis of the gag gene products, Pr76gag and Pr180gag-pol, which was the precursor of active reverse transcriptase. Thus the HNP produced virions that were not infectious due to a defect of the env gene. Studies on the viral RNA showed that a 35 S RNA, estimated to be 8000 nucleotides long, was the genomic RNA of AEV-H and probably carried one transforming gene, which is most likely erbB gene. The gene organization of AEV-H was suggested to be 5'-gag-pol-onc-3'. These data imply that the single oncogene is responsible for both erythroblastosis and sarcoma.

A 27-year-old man developed bilateral destructive ocular manifestations of generalized Wegener's granulomatosis. Before the appearance of necrotic granuloma, which replaced almost all the original ocular components, the patient had suffered from severe headache and restricted ocular motility for five years. A general remission was achieved with azathioprine and prednisolone treatment although the patient is blind because of destruction of the globes.

Antigen receptors on the T-cell surface are noncovalently associated with at least four invariant polypeptide chains, CD3-gamma, -delta, -epsilon, and -zeta. The mouse CD3-gamma gene, consisting of seven exons, was found to be highly homologous to the CD3-delta gene described earlier. Both the high level of sequence homology and the exon/intron organization indicate that the CD3-gamma and -delta genes arose by gene duplication. Surprisingly, murine and human genomic DNA clones could be isolated that contained elements of both the CD3-gamma and CD3-delta genes. In fact, the putative transcription start site of the mouse CD3-gamma gene is less than 1.4 kilobases from the transcription initiation site of the mouse CD3-delta gene. Common elements that regulate the divergent transcription of the two genes are therefore proposed to be located in the intervening 1.4-kilobase DNA segment. This might contribute to the coordinate expression of the CD3-gamma and -delta genes during intrathymic maturation of T lymphocytes.

Using a subtractive cDNA cloning method, we isolated a number of T-lymphocyte-specific genes. One of these genes, represented by the cDNA clone pT49, is expressed in cytotoxic T lymphocytes but not in helper T lymphocytes or B lymphocytes. The protein structure deduced from the nucleotide sequence showed a high degree of homology to fibrinogen beta and gamma subunits. This might indicate that evolutionarily fibrinogen has its origin in lymphocytes. In spite of the strong homology of pT49 protein to the fibrinogen subunits, the positions of the introns in the pT49 gene are totally different from those of the fibrinogen genes.