An Indirect Cost of Male-Male Aggression Arising from Female Response.

Animal behavior is often polymorphic between individuals within a population. A cost/benefit balance of a particular behavioral pattern may be influenced by social interaction with other individuals with different behavioral patterns. Males of a fruitfly, Drosophila prolongata, show genetically defined polymorphism in aggressiveness and boldness against rival males. Males of the H strain are highly aggressive, and their fights tend to escalate into boxing, the highest level of aggressive interaction. H males are also bold against sneaker males and do not hesitate to perform leg vibration (LV), a courtship behavior that is vulnerable to interception of the female by surrounding rival males. In contrast, males of the L strain rarely engage in boxing and do not perform LV in the presence of rival males. We examined their mating success in small experimental populations. The mating success of L males was higher in a pure L population than in a mixed population with H males, whereas that of H males was higher in a mixed population than in a pure H population. Notably, this 'cost of aggression' in a pure H population seemed not directly derived from the male-to-male interaction but was imposed by the female's response of escaping from fighting males, compromising the benefit of the resource monopolization as territory.

[Effective Treatment of Ameloblastoma in an Odontogenic Cyst with Fenestration before Curative Surgery-A Case Report].

Ameloblastoma is a benign odontogenic tumor that rarely undergoes malignant transformation and metastasis but may be locally invasive and recurrent. Fenestration is used to reduce maxillary odontogenic cysts. Here, we report a case ofameloblastoma that developed in the wall of an odontogenic cyst and was treated with fenestration before curative surgery. A 57-yearold Japanese man presented with a mass on the right side ofthe lower gingiva. Computed tomography revealed a unicystic lesion in the right mandibular body, accompanied by a multicystic area in the right lower canine region. Three involved molars were extracted and the cystic wall was harvested. Biopsy analysis revealed an odontogenic cyst in the unicystic lesion and an ameloblastoma in the multicystic area. The ameloblastoma was thought to have developed in the odontogenic cyst wall. The biopsy wound was maintained as a fenestration for 3 months and the lesion was reduced. Marginal resection of the mandible with cystectomy was performed to preserve mandibular bone continuity and the mandibular nerve. Although fenestration delayed curative surgery, the large cystic lesion reduction helped to avoid complications after curative surgery.

Identification of a Direct Biosynthetic Pathway for UDP-N-Acetylgalactosamine from Glucosamine-6-Phosphate in Thermophilic Crenarchaeon Sulfolobus tokodaii.

Most organisms, from Bacteria to Eukarya, synthesize UDP-N-acetylglucosamine (UDP-GlcNAc) from fructose-6-phosphate via a four-step reaction, and UDP-N-acetylgalactosamine (UDP-GalNAc) can only be synthesized from UDP-GlcNAc by UDP-GlcNAc 4-epimerase. In Archaea, the bacterial-type UDP-GlcNAc biosynthetic pathway was reported for Methanococcales. However, the complete biosynthetic pathways for UDP-GlcNAc and UDP-GalNAc present in one archaeal species are unidentified. Previous experimental analyses on enzymatic activities of the ST0452 protein, identified from the thermophilic crenarchaeon Sulfolobus tokodaii, predicted the presence of both a bacterial-type UDP-GlcNAc and an independent UDP-GalNAc biosynthetic pathway in this archaeon. In the present work, functional analyses revealed that the recombinant ST2186 protein possessed an glutamine:fructose-6-phosphate amidotransferase activity and that the recombinant ST0242 protein possessed a phosphoglucosamine-mutase activity. Along with the acetyltransferase and uridyltransferase activities of the ST0452 protein, the activities of the ST2186 and ST0242 proteins confirmed the presence of a bacterial-type UDP-GlcNAc biosynthetic pathway in S. tokodaii In contrast, the UDP-GlcNAc 4-epimerase homologue gene was not detected within the genomic data. Thus, it was expected that galactosamine-1-phosphate or galactosamine-6-phosphate (GalN-6-P) was provided by conversion of glucosamine-1-phosphate or glucosamine-6-phosphate (GlcN-6-P). A novel epimerase converting GlcN-6-P to GalN-6-P was detected in a cell extract of S. tokodaii, and the N-terminal sequence of the purified protein indicated that the novel epimerase was encoded by the ST2245 gene. Along with the ST0242 phosphogalactosamine-mutase activity, this observation confirmed the presence of a novel UDP-GalNAc biosynthetic pathway from GlcN-6-P in S. tokodaii Discovery of the novel pathway provides a new insight into the evolution of nucleotide sugar metabolic pathways.IMPORTANCE In this work, a novel protein capable of directly converting glucosamine-6-phosphate to galactosamine-6-phosphate was successfully purified from a cell extract of the thermophilic crenarchaeon Sulfolobus tokodaii Confirmation of this novel activity using the recombinant protein indicates that S. tokodaii possesses a novel UDP-GalNAc biosynthetic pathway derived from glucosamine-6-phosphate. The distributions of this and related genes indicate the presence of three different types of UDP-GalNAc biosynthetic pathways: a direct pathway using a novel enzyme and two conversion pathways from UDP-GlcNAc using known enzymes. Additionally, Crenarchaeota species lacking all three pathways were found, predicting the presence of one more unknown pathway. Identification of these novel proteins and pathways provides important insights into the evolution of nucleotide sugar biosynthesis, as well as being potentially important industrially.

[A Case of Medical Leech Therapy for Venous Congestion Following Forearm Flap Reconstruction].

Leeches have been used for medical treatment for at least 2,500 years. Plastic surgeons have recently begun to use leeches to reduce venous congestion after flap reconstruction. However, few reports of leech use in the oral region have been published. We report a case of medical leech therapy used to treat venous congestion after forearm flap reconstruction for oral cancer. A 67-year-old female was diagnosed with squamous cell carcinoma of the left tongue margin(cT2N0M0, Stage Ⅱ). The patient underwent tracheostomy, supraomohyoid neck dissection, hemiglossectomy, and reconstruction using a free forearm flap under general anesthesia. Venous congestion in the forearm flap was detected 21 hours postoperatively, and reanastomosis of the flap was performed. However, venous congestion continued after revision surgery. Therefore, we introduced medical leech therapy to treat the venous congestion. Leeches were used twice daily for 5 days, and the total hematophagy volume was 21.6 g. After leech treatment, continuous bleeding from the skin flap decreased and skin color improved. The medial skin flap survived, and the patient was able to eat 13 days after the initial operation. The rest of the treatment has been uneventful to date without dysfunction of the skin flap.

[A Contrivance for Closure and Dressing of Orocutaneous Fistula Developed in Advanced Oral Cancer].

Orocutaneous fistula sometimes occurs in locallyadvanced unresectable or recurrent oral squamous cell carcinoma. The developed orocutaneous fistula results in constant leakage of saliva, ingested foods and liquids and decline in patients' quality of life(QOL). A 47-year-old Japanese man had received treatment for tongue carcinoma. At the routine follow-up, a cystic lesion in the right submandibular region was detected. Biopsyof the specimen of the cystic lesion revealed squamous cell carcinoma. After chemotherapy, an orocutaneous fistula between the right oropharyngeal and the right submandibular region developed and graduallyincreased. Although closure and dressing of the orocutaneous fistula with various materials was attempted, it was ultimatelyunsuccessful. Finally, application of a rubber film and silicone adhesive agent to the skin was successful for closure and dressing of the fistula. Orocutaneous fistula is one of major contributors to decline in patients' QOL. The sharing of information regarding effective methods or materials for closure and dressing of orocutaneous fistula is necessaryto maintain patients' QOL.

Identification and characterization of a thermostable bifunctional enzyme with phosphomannose isomerase and sugar-1-phosphate nucleotidylyltransferase activities from a hyperthermophilic archaeon, Pyrococcus horikoshii OT3.

Mannosylglycerate is known as a compatible solute, and plays important roles for salinity adaptation and high temperature stability of microorganisms. In the gene cluster for the mannosylglycerate biosynthetic pathway predicted from the genomic data of Pyrococcus horikoshii OT3, the PH0925 protein was found as a putative bifunctional enzyme with phosphomannose isomerase (PMI) and mannose-1-phosphate guanylyltransferase (Man-1-P GTase) activities, which can synthesize GDP-mannose when accompanied by a phosphomannomutase/phosphoglucomutase (PMM/PGM) enzyme (PH0923). The recombinant PH0925 protein, expressed in E. coli, exhibited both expected PMI and Man-1-P GTase activities, as well as absolute thermostability; 95 °C was the optimum reaction temperature. According to the guanylyltransferase activity (GTase) of the PH0925 protein, it was found that the protein can catalyze glucose-1-phosphate (Glc-1-P) and glucosamine-1-phosphate (GlcN-1-P) in addition to Man-1-P. The analyses of C-terminus-truncated forms of the PH0925 protein indicated that sugar-1-phosphate nucleotidylyltransferase (Sugar-1-P NTase) activity was located in the region from the N-terminus to the 345th residue, and that the C-terminal 114 residue region of the PH0925 protein inhibited the Man-1-P GTase activity. Conversely, the PMI activity was abolished by deletion of the C-terminal 14 residues. This is the first report of a thermostable enzyme with both PMI and multiple Sugar-1-P NTase activities.

Drosophila suzukii preferentially lays eggs on spherical surfaces with a smaller radius.

Drosophila suzukii is an agricultural pest that predominantly harms small fruits, having a serrated ovipositor that is able to pierce the skin of ripening fruits. Its oviposition preference has been studied from various aspects including chemical and physical properties of oviposition substrates. However, its preference for certain shapes or sizes of substrates has not been explored. In this study, we tested the oviposition preference of D. suzukii for artificial oviposition substrates with different surface curvatures using 27 strains recently established from wild populations collected in Japan. We found that D. suzukii laid more eggs on a surface with smaller radii (4.8 and 5.7 mm) compared with larger radii (7.7 and 9.6 mm). We also found that the most preferred radius differed among strains. Notably, the preference was independent of the volume of substrates, suggesting that D. suzukii uses the surface curvature as a cue for its oviposition site selection. These results provide an additional explanation for why D. suzukii preferentially uses small fruits as its oviposition sites.

Fenestration is a logical and effective treatment for a large primordial cyst with cholesterol granuloma: a case report.

A cholesterol granuloma (CG) is characterized by the presence of cholesterol crystals that cause a chronic granulomatous reaction in an enclosed space. It occurs most commonly in the head and neck region, particularly in the middle ear. Although CGs in the maxilla have also been reported, odontogenic cysts in conjunction with CGs in the maxilla are very rare. We herein present a case of a 72-year-old man who developed a large primordial cyst with a maxillary CG that extended into the maxillary sinus, nasal cavity, and infraorbital region, causing left-sided facial swelling and discomfort. We successfully controlled the symptoms and reduced the size of the lesion using the treatment approach for a common odontogenic cyst: fenestration followed by complete excision. This case suggests that fenestration is an effective technique to treat odontogenic cysts with CGs. Although the mechanisms underlying the pathogenesis and growth of CGs are still unknown, our report highlights a potential therapeutic approach for these lesions.

Identification of novel acetyltransferase activity on the thermostable protein ST0452 from Sulfolobus tokodaii strain 7.

A 401-residue-long protein, ST0452, has been identified from a thermophilic archaeon, Sulfolobus tokodaii strain 7, as a glucose-1-phosphate thymidylyltransferase (Glc-1-P TTase) homolog with a 170-residue-long extra C-terminus portion. Functional analyses of the ST0452 protein have confirmed that the protein possessed dual sugar-1-phosphate nucleotidylyltransferase (sugar-1-P NTase) activities. The 24 repeats of a signature motif sequence which has been found in bacterial acetyltransferases, (L/I/V)-(G/A/E/D)-XX-(S/T/A/V)-X, were detected at the C terminus of the ST0452 protein. This observation prompted our group to investigate the acetyltransferase activity of the ST0452 protein. Detection of the release of coenzyme A (CoA) from acetyl-CoA and the production of UDP-N-acetyl-d-glucosamine (UDP-GlcNAc) from glucosamine-1-phosphate (GlcN-1-P) and UTP in the presence of the ST0452 protein revealed that this protein possesses the GlcN-1-P-specific acetyltransferase activity. In addition, analyses of substrate specificity showed that acetyltransferase activity of the ST0452 protein is capable of catalyzing the change of galactosamine-1-phosphate (GalN-1-P) to N-acetyl-d-galactosamine-1-phosphate (GalNAc-1-P) as well as GlcN-1-P and that its sugar-1-P NTase activity is capable of producing UDP-GalNAc from GalNAc-1-P and UTP. This is the first report of a thermostable bifunctional enzyme with GalN-1-P acetyltransferase and GalNAc-1-P uridyltransferase activities. The observation reveals that the bacteria-type UDP-GlcNAc biosynthetic pathway from fructose-6-phospate is utilized in this archaeon and represents a novel biosynthetic pathway for producing UDP-GalNAc from GalN-1-P in this microorganism.

Fracture of the Clavicle following Radical Neck Dissection and Reconstruction Using Pectoralis Major Myocutaneous Flap Accompanied by Postoperative Radiotherapy.

Fracture of the clavicle following radical neck dissection (RND) and/or radiotherapy is a rare complication. Several causes of fracture of the clavicle after treatment of head and neck cancer were postulated in previous reports. We present a case of fracture of the clavicle after treatment of squamous cell carcinoma of the tongue. An 81-year-old Japanese woman underwent RND, subtotal glossectomy, reconstruction using a pectoralis major myocutaneous flap (PMMCF), and postoperative radiotherapy (50.4 Gy). One month after the primary treatment, fracture of the clavicle occurred. It was thought that muscular dynamic factor and reduction of blood supply in the clavicle associated with RND and PMMCF were the causes of the fracture. We have to recognize the occurrence of this complication and try to reduce the factors related to the complication.

Increasing in archaeal GlcNAc-1-P uridyltransferase activity by targeted mutagenesis while retaining its extreme thermostability.

UDP-GlcNAc, an activated and essential form of GlcNAc which is an important component in the polysaccharide structure of most organisms, is synthesized from GlcNAc-1-P and UTP by GlcNAc-1-P UTase. We previously reported the identification of the extremely thermostable ST0452 protein, which has dual sugar-1-P NTase activities (Glc-1-P TTase and GlcNAc-1-P UTase activities) from an acidothermophilic archaeon, Sulfolobus tokodaii strain 7. Detailed analyses of the protein indicated that the activity is slightly lower than that of bacteria. For industrial applications, activity needs to be increased without decreasing thermostability. Therefore, to enhance this activity, we introduced mutations into the amino acid residues located within the predicted reaction centre by targeted mutagenesis. All 12 mutant ST0452 proteins showed no decrease in thermostability. Among them, six mutant proteins were found to have increased GlcNAc-1-P UTase activity under optimal reaction conditions with sufficient substrates or an appropriate metal ion. Our results indicate that targeted mutagenesis is a powerful technique for in vitro production of a thermostable enzyme with enhanced activity. The results of this study also indicate that the space for the metal ion is important for selecting the type of metal ion and also affects the rate of the reaction.

Characterization of a thermostable enzyme with phosphomannomutase/phosphoglucomutase activities from the hyperthermophilic archaeon Pyrococcus horikoshii OT3.

The phosphomannomutase/phosphoglucomutase (PMM/PGM) enzyme catalyzes reversibly the intra-molecular phosphoryl interconverting reaction of mannose-6-phosphate and mannose-1-phosphate or glucose-6-phosphate and glucose-1-phosphate. Glucose-6-phosphate and glucose-1-phosphate are known to be utilized for energy metabolism and cell surface construction, respectively. PMM/PGM has been isolated from many microorganisms. By performing similarity searches using existing PMM/PGM sequences, the homologous ORFs PH0923 and PH1210 were identified from the genomic data of Pyrococcus horikoshii OT3. Since PH0923 appears to be part of an operon consisting of four carbohydrate metabolic enzymes, PH0923 was selected as the first target for the investigation of PMM/PGM activity in P. horikoshii OT3. The coding region of PH0923 was cloned and the purified recombinant protein was utilized for an examination of its biochemical properties. The enzyme retained half its initial activity after treatment at 95 degrees C for 90 min. Detailed analyses of activities showed that this protein is capable of utilizing a variety of metal ions that are not utilized by previously characterized PMM/PGM proteins. A mutated protein with an alanine residue replacing the active site serine residue indicated that this residue plays an important but non-essential role in PMM/PGM activity.

Identification of an extremely thermostable enzyme with dual sugar-1-phosphate nucleotidylyltransferase activities from an acidothermophilic archaeon, Sulfolobus tokodaii strain 7.

L-rhamnose is an essential component of the cell wall and plays roles in mediating virulence and adhesion to host tissues in many microorganisms. Glucose-1-phosphate thymidylyltransferase (RmlA, EC 2.7.7.24) catalyzes the first reaction of the four-step pathway of L-rhamnose biosynthesis, producing dTDP-D-glucose from dTTP and glucose-1-phosphate. Three RmlA homologues of varying size have been identified in the genome of a thermophilic archaeon, Sulfolobus tokodaii strain 7. In this study, we report the heterologous expression of the largest homologue (a 401 residue-long ST0452 protein) and characterization of its thermostable activity. RmlA enzymatic activity of this protein was detected from 65 to 100 degrees C, with a half-life of 60 min at 95 degrees C and 180 min at 80 degrees C. Analysis of a deletion mutant lacking the 170-residue C-terminal domain indicated that this region has an important role in the thermostability and activity of the protein. Analyses of substrate specificity indicated that the enzymatic activity of the full-length protein is capable of utilizing alpha-D-glucose-1-phosphate and N-acetyl-D-glucosamine-1-phosphate but not alpha-D-glucosamine-1-phosphate. However, the protein is capable of utilizing all four deoxyribonucleoside triphosphates and UTP. Thus, the ST0452 protein is an enzyme containing both glucose-1-phosphate thymidylyltransferase and N-acetyl-D-glucosamine-1-phosphate uridylyltransferase activities. This is the first report of a thermostable enzyme with dual sugar-1-phosphate nucleotidylyltransferase activities.

Development of an integrated automation system with a magnetic bead-mediated nucleic acid purification device for genetic analysis and gene manipulation.

We have developed an integrated automation system for genetic analysis and gene manipulation. The system, SX-8G Plus, is equipped with an 8-nozzle dispensing unit, a thermal cycler, a cooled reagent reservoir, four tip storage racks, four microplate platforms, buffer reservoirs, an agarose gel electrophoresis unit, a power supply, a pump for exchanging electrophoresis buffer, and a CCD camera. Automation of nucleic acid extraction and purification, the most difficult step in automating genetic analysis and gene manipulation, was realized using magnetic beads with Magtration Technology, which we have previously developed for automating the handling of paramagnetic beads. Using this system, we could perform the automated separation and purification of DNA fragments by agarose gel electrophoresis starting from sample loading. The system would enable the automation of almost all procedures in genetic analysis and gene manipulation.