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Harmful algal blooms caused by raphidophyte species of the genus Chattonella (i.e. Chattonella antiqua, Chattonella marina and Chattonella ovata) have been documented in temperate coastal regions around the world. To understand the effects of physicochemical factors on bloom development of Chattonella spp., we investigated the variations of vegetative and resting cells (i.e. cysts) of Chattonella spp. and environmental variables in two coastal environments, Uranouchi Inlet (extremely closed) and Nomi Inlet (semi-closed), with contrasting enclosed natures. Although the vegetative cells and cysts of Chattonella spp. were distributed in both coastal regions, the densities were remarkably higher in Uranouchi Inlet than in Nomi Inlet. The mud content in the sediments of Uranouchi Inlet was also higher than that in the sediments of Nomi Inlet, meaning that fine particles such as cysts are likely to accumulate in the former region. Because of the extremely closed nature of Uranouchi Inlet, warm oceanic waters of the Kuroshio Current penetrate the inlet only infrequently. These results suggest that the closed nature of coastal regions is an important factor influencing either water exchange or the resultant accumulation of Chattonella cells in coastal environments.

Benthic dinoflagellates of the genus Ostreopsis are found all over the world in temperate, subtropical, and tropical coastal regions. Our recent studies revealed that a putative “cryptic” species of Ostreopsis ovata is present widely along Japanese coasts. This organism, Ostreopsis sp. 1, possesses palytoxin analogs and thus its toxic blooms may be responsible for potential toxification of marine organisms. To evaluate the bloom dynamics of Ostreopsis sp. 1, the present study examined the growth responses of Ostreopsis sp. 1 strain s0716 to various light intensities (photon flux densities: μmol photons m −2 s −1 ) using a newly devised photoirradiation-culture system. This novel system has white light-emitting diodes (LEDs) capable of more closely simulating the wavelength spectrum of light entering the oceanic water column than do fluorescent tubes and halogen lamps. In this system, the light intensity of the white LEDs was reduced through two polarizing filters by varying the rotation angles of the filters. Thereby, the new system was capable of culturing microalgae under well-controlled light intensity conditions. Ostreopsis sp. 1 grew proportionally when light intensity was increased from 49.5 to 199 μmol photons m −2 s −1 , but its growth appeared to be inhibited slightly at ≥263 μmol photons m −2 s −1 . The relationship between observed growth rates and light intensity was calculated at R > 0.99 ( P Ostreopsis sp. 1 and the organism's growth potential as follows: (1) the threshold light intensity for growth: 29.8 μmol photons m −2 s −1 ; (2) the optimum light intensity ( L m ) giving the maximum growth rate ( μ max = 0.659 divisions day −1 ): 196 μmol photons m −2 s −1 ; (3) the optimum light intensity range ( L opt ) giving ≥95% μ max : 130–330 μmol photons m −2 s −1 ; (4) the semi-optimum range ( L sopt ) giving ≥80% μ max : 90 to over 460 μmol photons m −2 s −1 . The L sopt represents 4.5–23% ambient light intensity present in surface waters off of a temperate region of the Japanese coast, Tosa Bay; putatively, this semi-optimum range of light intensity appears at depth of 12.9–27.8 m. Considering these issues, our data indicate that Ostreopsis sp. 1 in coastal environments may form blooms at ca. ∼28 m depth in regions along Japanese coasts.

Summary Benthic dinoflagellates of the genus Ostreopsis produce palytoxin (PTX)-like compounds. The worldwide distributed Ostreopsis ovata/O. cf. ovata is potentially responsible for outbreaks of human health problems around the coasts of tropical, subtropical, and temperate regions. The present study examined growth responses of an O. cf. ovata strain s0662 collected from coastal waters of Japan with 35 different combinations of temperature (15–35°C) and salinity (20–40) and discusses the bloom dynamics of the organism in Japanese coastal environments. The O. cf. ovata strain s0662 tolerated a wide range of temperature (17.5–35°C) and salinity (25–40). Results of a two-way ANOVA showed significant effects of temperature-salinity interaction on growth rates and biomass yields of the O. cf. ovata strain (F(24,70) > 127, P

Blooms of toxic dinoflagellates of the genus Ostreopsis, which is known as a producer of palytoxin (PTX) analogs, may pose a threat to human health in tropical, subtropical, and temperate regions around the world. In the present study, we established a suitable culture method for Ostreopsis spp. isolated from Japanese coastal waters and characterized their growth potential using the method to discuss their bloom dynamics. Each clonal strain of Ostreopsis cf. ovata, Ostreopsis sp. 1, Ostreopsis sp. 5, and Ostreopsis sp. 6 was incubated in 25 × 150 mm test tubes with a flat bottom containing various kinds of medium. Since Ostreopsis spp. strains grew well in IMK and/or f/2 media, we selected these media for cultivation of all the Ostreopsis spp. isolates. Growth rates of O. cf. ovata (0.834 divisions/day), Ostreopsis sp. 1 (0.619 divisions/day), and Ostreopsis sp. 6 (1.04 divisions/day) that produce PTX analogs significantly differed (p < 0.05) respectively and are clearly higher than those of other reported epiphytic dinoflagellate Gambierdiscus toxicus, Prorocentrum lima, and Coolia monotis cultures, which suggest that these species have ecological advantages to predominate through the algal succession in Japanese coastal waters, resulting in a potential risk to human health in this region.

This study evaluates the craniofacial change during the third decade of life and compared the changes that occurred in subjects with normal occlusion and untreated malocclusions. 10 cases each of normal occlusion, untreated maxillary protrusion and mandibular protrusion were subjected to cephalometric analysis and model analysis. The mean age at the beginning of the observation (T1) was 24 years, and the observation period (T2) was approximately 13 years in three groups. There was no significant difference in any cephalometric values between T1 and T2 in the normal occlusion group. The SNB angle was slightly decreased, the Y-axis was increased, and upper incisors were lingually inclined in the maxillary protrusion group and increasing of overjet and decreasing of overbite was observed in the mandibular protrusion group. In model analysis, the arch length discrepancy (ALD) and the inter-molar width (IMW) were slightly increased in the lower arch in the normal occlusion group and the upper arch length (AL) had decreased in the mandibular protrusion group. In conclusion, significant differences were found in the changes of several skeletal and dental parameters between normal occlusion and untreated malocclusion subjects in the third decade of life. These changes in the third decade of life should be considered in order to plan the orthodontic treatment and evaluate the long-term stability in the patients with maxillary and mandibular protrusion.

Several species of the genus Karenia (Dinophyceae) form blooms and often cause the mortality of cultured and wild fish. In Japan, blooms caused by two species - namely Karenia mikimotoi and Karenia brevis - have been reported so far. On the basis of morphological and molecular-phylogenic examinations, the present investigation found Karenia papilionacea and its novel sister phylotype for the first time in the coastal waters of the various regions of Japan. Of 34 strains isolated from the coastal waters, 27 strains displayed the typical morphological characteristics of K. papilionacea and further showed consensus DNA sequences corresponding to those of the originally described K. papilionacea. The other 7 strains displayed the same morphological characteristics of K. papilionacea, but showed divergent DNA sequences, at a genetic distance of over 0.04 (Internal Transcribed Spacer regions) from those of the original phylotype of K. papilionacea. These divergent strains were characterized as a novel sister phylotype (phylotype-I) of K. papilionacea. In the coastal waters of Japan, K. papilionacea-like (K. papilionacea and/or its phylotype-I) formed blooms at 20.3-30.4°C and salinity 30.1-33.9. No K. brevis was identified in Japanese coastal waters during this study. These findings demonstrated that K. papilionacea occurs along the coasts of western Japan and possibly shares several coastal regions with K. mikimotoi and K. papilionacea phylotype-I. In order to assess the risks of Karenia blooms to aquaculture, it is essential that the growth physiology and ichthyotoxicity of K. papilionacea and its novel phylotype should be characterized.

Patients with open bite often show a weak occlusal force and temporomandibular disorders (TMDs). If these are the main cause of open bite, it may be hypothesized that both pre-pubertal and adult open-bite patients would show a weak occlusal force and abnormal condylar motion. The purpose of this study was to test this hypothesis. Test group subjects consisted of 13 consecutive pre-pubertal and 13 adult patients with anterior open bite. They were compared with age-matched normal subjects. The adult open-bite group showed a weaker occlusal force and a shorter range of condylar motion compared with the control subjects. In the pre-pubertal subjects, however, there were no significant differences in the occlusal force and range of condylar motion between the open-bite and control groups. Therefore, these results suggest that a weak occlusal force or TMDs may not be the main cause of open bite.

The purpose of this study was to test the hypothesis that the application of occlusal splints increases the diurnal salivary flow rate both in bruxism patients and in normal subjects. Salivary flow rates in 16 adult volunteers (8 bruxism patients and 8 sex- and age-matched control subjects) were measured with the spitting method. There was no significant difference in the salivary flow rate with or without splints between the control and bruxism groups. In all subjects, the salivary flow rates with splints were significantly higher than those without splints during relaxing, clenching, and chewing-like movement. The salivary flow rate during the chewing-like movement was significantly higher than that during relaxing and clenching, irrespective of splint application. The results suggest that maxillary occlusal splints might stimulate salivary secretion, particularly during chewing-like movement, in both bruxism patients and normal subjects.

Study objective To examine the relationship between nocturnal bruxism and gastroesophageal reflux. Design Controlled descriptive study and double-blind, placebo-controlled, clinical study. Setting Portable pH monitoring, electromyography, and audio-video recordings were conducted during the night in the subjects' home. Participants Ten patients with bruxism and 10 normal subjects were matched for height, weight, age, and sex. They did not have symptoms of gastroesophageal reflux disease. Intervention Medication with a proton pump inhibitor (ie, a gastric-acid-inhibiting drug). Measurements and results The bruxism group showed a significantly higher frequency of nocturnal rhythmic masticatory muscle activity (RMMA) episodes (mean +/- SD: 6.7 +/- 2.2 times per hour) and a higher frequency and percentage of time of gastroesophageal reflux episodes with a pH less than 4.0 and 5.0 (0.5 +/- 0.9 and 3.6 +/- 1.6 times per hour and 1.3% +/- 2.5% and 7.4% +/- 12.6%, respectively) than the control group (RMMA episodes: 2.4 +/- 0.9 times per hour; gastroesophageal reflux episodes: 0.0 +/- 0.0 and 0.1 +/- 0.3 times per hour and 0.0% +/- 0.0% and 0.0% +/- 0.0%, respectively). In the bruxism group, 100% of the gastroesophageal reflux episodes with a pH less than 3.0 and 4.0 included both an RMMA episode and an electromyographic burst, the duration of which was approximately 0.5 to 1.0 seconds, probably representing swallowing of saliva. The majority of gastroesophageal reflux episodes with a pH of 4.0 to 5.0 also included both an RMMA episode and an electromyographic burst in the control and bruxism groups (100% +/- 0.0% vs 70.7% +/- 16.5%), again probably due to swallowing of saliva. The remaining minority of gastroesophageal reflux episodes with a pH of 4.0 to 5.0 contained only an electromyographic burst (swallowing of saliva). The frequency of RMMA episodes after the release of the medication from the proton pump inhibitor, which increased the gastric and esophageal pH, was significantly lower than that after administration of the placebo in the control and bruxism groups (1.0 +/- 0.6 vs 1.9 +/- 3.2 times per hour, and 3.7 +/- 1.9 vs. 6.0 +/- 2.2 times per hour, respectively). Conclusions Nocturnal bruxism may be secondary to nocturnal gastroesophageal reflux, occurring via sleep arousal and often together with swallowing. The physiologic link between bruxism and the increase in salivation needs to be investigated.

The influence of reduced anterior disc displacement on condylar motion has not been fully examined in young adults. Reduced anterior disc displacement was hypothesized to inhibit condylar motion. Using a six-degrees-of-freedom jaw-tracking system, we recorded bilateral condylar motion during maximum open-close jaw movement and gum-chewing on both sides in ten young adults with unilateral reduced anterior disc displacement and in ten control subjects without temporomandibular disorders. The bilateral condylar motion during both maximum open-close jaw movement and chewing on the disc-displacement side was inhibited in the test group. The condylar motion on the disc-displacement side during chewing on the non-disc-displacement side was also inhibited in the test group. These results suggest that the limitation of condylar motion on the disc-displacement side may influence condylar motion on the non-disc-displacement side during maximum open-close jaw movement, and mastication on the disc-displacement side in young adults.

It has been reported that loading to the mandible during closing movement makes the condylar path move more in the superior direction than that during the free closing movement. In this study, the hypothesis was tested that the displacement of the condyle on the chewing side is greater in the direction of the mandibular fossa than that on the non-chewing side. Using a six-degrees-of-freedom jaw movement recording system, we recorded condylar motion in 12 healthy adults without TMD, during the chewing of a large hard gummy jelly. The maximum displacements at the condyle on the chewing side from the maximum intercuspation (CO) position were significantly larger in the superior and medial directions at the initial stage and in the posterior direction at all stages (0.5 mm, 0.5 mm, and 0.6 mm, respectively) than those on the non-chewing side (0.0 mm, 0.1 mm, and 0.1 mm, respectively). This suggests that, in healthy adults, the condyles at CO are located in a position such that excessive load is not applied to the temporomandibular joint when there are the aforementioned displacements.

Mesophilic cytochrome c551 of Pseudomonas aeruginosa (PA c551) became as stable as its thermophilic counterpart, Hydrogenobacter thermophilus cytochrome c552 (HT c552), through only five amino acid substitutions. The five residues, distributed in three spatially separated regions, were selected and mutated with reference to the corresponding residues in HT c552 through careful structure comparison. Thermodynamic analysis indicated that the stability of the quintuple mutant of PA c551 could be partly attained through an enthalpic factor. The solution structure of the mutant showed that, as in HT c552, there were tighter side chain packings in the mutated regions. Furthermore, the mutant had an increased total accessible surface area, resulting in great negative hydration free energy. Our results provide a novel example of protein stabilization in that limited amino acid substitutions can confer the overall stability of a natural highly thermophilic protein upon a mesophilic molecule., This work was supported by a grant from the Japanese Ministry of Education, Science and Culture.

Purpose The purpose of this study was to examine the correlation between the condylar path length and craniofacial morphology in patients with mandibular prognathism and deviation, and investigate changes in the condylar path length and temporomandibular joint disorders after intraoral vertical ramus osteotomy (IVRO). Patients and Methods Sixteen Japanese patients exhibiting mandibular prognathism and deviation were studied. For their correction, all patients underwent IVRO. In the case of patients with severe occlusal cant after cant correction by Le Fort I osteotomy, IVRO was carried out on the same day. The change in craniofacial morphology was assessed with frontal cephalograms and submentovertex radiograms. Condylar path changes were assessed using a 6° of freedom measuring device. All patients were examined with regard to their temporomandibular joint (TMJ) function. Results Before treatment, mandibular deviation was moderately related to the asymmetries of craniofacial morphology and condylar path length. The rate of incidence of TMJ sounds on the deviated side was significantly higher than that on the nondeviated side. After treatment, although the condylar path length as well as condylar position had changed to become bilaterally symmetric, there was still a significant difference during protrusive excursion and maximal open–close movements. TMJ sounds disappeared in 64.3% of patients. Conclusion Mandibular deviation was found to be strongly related to the morphologic and functional asymmetries in patients with mandibular prognathism and deviation. The condylar path length and condylar position were improved after the correction of mandibular deviation with IVRO.

An eight-year-, four-month-old girl was brought to the orthodontic clinic of Okayama University Medical and Dental Hospital. The patient had an impacted upper left second premolar because of an odontogenic keratocyst and showed a skeletal Class II jaw base relationship. At the age of six years four months, marsupialization of a cyst was performed at the Okayama University Medical and Dental Hospital because the patient had shown a swelling of the left cheek because of the cyst. The upper left second premolar was located in the roof of the maxillary sinus. The cyst was histopathologically diagnosed as an odontogenic keratocyst. At the age of nine years 10 months and after regaining the space for eruption of the premolar, the impacted premolar erupted without traction. At the age of 12 years five months, edgewise treatment was initiated, which continued for three years. After removing the edgewise appliance, an optimum occlusion was achieved. The occlusion was maintained without recurrence of the keratocyst after a retention period of five years.

The purpose of this study was to examine the relationships among nocturnal jaw muscle activities, decreased esophageal pH, and sleep positions. Twelve adult volunteers, including 4 bruxism patients, participated in this study. Portable pH monitoring, electromyography of the temporal muscle, and audio-video recordings were conducted during the night in the subjects' homes. Rhythmic masticatory muscle activity (RMMA) episodes were observed most frequently, with single short-burst episodes the second most frequent. The frequencies of RMMA, single short-burst, and clenching episodes were significantly higher during decreased esophageal pH episodes than those during other times. Both the electromyography and the decreased esophageal pH episodes were most frequently observed in the supine position. These results suggest that most jaw muscle activities, ie, RMMA, single short-burst, and clenching episodes, occur in relation to gastroesophageal reflux mainly in the supine position.

Patients with unilateral posterior crossbite often show reverse sequential jaw movement patterns on the frontal view during mastication on the crossbite side. Recent studies show that such patients are prone to suffer from temporomandibular joint (TMJ) disc displacement, particularly the lateral portion. The purpose of this study was to examine the movement of the lateral and medial poles of the working condyle during mastication in such patients. Subjects were 12 consecutive patients with unilateral posterior crossbites and without TMJ disc displacements and 12 normal subjects. An optoelectronic jaw-tracking system with 6 degrees of freedom was used to record the motion of the lateral and medial poles of the working condyle during mastication of standardized hard, gummy jelly. The data from the first 10 cycles were analyzed. The lateral and medial poles of the condyle on the crossbite side moved more in the medial direction and less in the lateral direction during mastication in the crossbite patients than the condyle in the normal subjects. The lateral pole of the working condyle moved more in the posterior and inferior directions and less in the anterior direction than the medial pole in all subjects. These results suggest that these condylar movements in patients with unilateral posterior crossbites might be related to the susceptibility to TMJ disc displacement, particularly the lateral portion.

Thermal stability was measured for variants of cytochrome c-551 (PA c-551) from a mesophile, Pseudomonas aeruginosa, and a thermophilic counterpart, Hydrogenobacter thermophilus cytochrome c-552 (HT c-552), by differential scanning calorimetry (DSC) at pH 3.6. The mutated residues in PA c-551, selected with reference to the corresponding residues in HT c-552, were located in three spatially separated regions: region I, Phe7 to Ala/Val13 to Met; region II, Glu34 to Tyr/Phe43 to Tyr; and region III, Val78 to Ile. The thermodynamic parameters determined indicated that the mutations in regions I and III caused enhanced stability through not only enthalpic but also entropic contributions, which reflected improved packing of the side chains. Meanwhile, the mutated region II made enthalpic contributions to the stability through electrostatic interactions. The obtained differences in the Gibbs free energy changes of unfolding [Delta(DeltaG)] showed that the three regions contributed to the overall stability in an additive manner. HT c-552 had the smallest heat capacity change (DeltaC(P)), resulting in higher DeltaG values over a wide temperature range (0-100 degrees C), compared to the PA c-551 variants; this contributed to the highest stability of HT c-552. Our DSC measurement results, in conjunction with mutagenesis and structural studies on the homologous mesophilic and thermophilic cytochromes c, provided an extended thermodynamic view of protein stabilization.

Although various antioxidants have been tested as therapeutics for endotoxemic subjects, the results of their efficacy are conflicting. Antioxidant biofactor (AOB) is a unique processed grain food that exhibits strong antioxidant activity (Minamiyama et al, J Nutr Sci Vitaminol, 40: 467-477, 1994). The present study was carried out to test the effect of AOB on hepatic injury in rats induced by lipopolysaccharide (LPS). Intravenous administration of LPS induced liver injury with a concomitant increase in hepatic generation of nitric oxide (NO) and 4-hydroxy-2-nonenal (HNE) modified proteins in the control group. The administration of AOB significantly inhibited the LPS-induced hepatic injury and generation of HNE-modified proteins and increased the survival rate of endotoxemic rats without affecting NO generation and plasma levels of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). AOB scavenged superoxide radicals without affecting NO production by LPS-stimulated macrophage cell line J 774.2 cells. AOB also inhibited lipid peroxidation induced by LPS in the cells. These results suggested that AOB might scavenge superoxide radicals and decrease toxic metabolites including HNE, thereby inhibiting liver injury in endotoxemic rats.