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Identifying target proteins that interact with bioactive molecules is indispensable for understanding their mechanisms of action. In this study, we developed a uniform ribosome display technology using equal-length DNAs and mRNAs to improve molecular display principle for target identification. The equal-length DNAs were designed to contain various coding sequences for full-length proteins with molecular weights of up to 130 kDa and were used to synthesize equal-length mRNAs, which allowed the formation of full-length protein-ribosome-equal-length mRNA complexes. Uniform ribosome display selections of dihydrofolate reductase and haloalkane dehalogenase mutant were performed against methotrexate and chlorohexane, respectively. Quantitative changes of proteins after each selection indicated that the target protein-displaying ribosomal complexes were specifically selected through non-covalent or covalent interactions with the corresponding bioactive molecules. Furthermore, selection of full-length proteins interacting with methotrexate or anti-DDX46 antibody from protein pools showed that only the target proteins could be precisely identified even though the molar amounts of equal-length mRNAs encoding them were adjusted to 1/20,000 of the total equal-length mRNAs. Thus, the uniform ribosome display technology enabled efficient identification of target proteins that interact with bioactive small and large molecules through simplified operations without deep sequencing.
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Three novel bacterial strains, FE4T, FE10T, and LA51T, which are phylogenetically affiliated to the genera Pseudoalteromonas, Vibrio, or Marinobacter, respectively, isolated from fertilized eggs and juveniles of sea cucumber Apostichopus japonicus were characterized by a genome-based taxonomical approach including multilocus sequence analysis (MLSA) combined with classical phenotypic and chemotaxonomic characterizations. A molecular network reconstructed on the basis of nucleotide sequences of four phylogenetic maker protein genes revealed that the strains FE4T, FE10T, and LA51T were closely related to Pseudoalteromonas shioyasakiensis, Vibrio lentus, and Marinobacter similis, respectively. Average nucleotide identity (ANI) comparisons against phylogenetically related species to FE4T, FE10T, and LA51T demonstrated that each newly described strain could not be identified as any previously described species within each genus showing < 95% ANI: 91.3% of FE4T against P. shioyasakiensis JCM 18891 T, 92.6% of FE10T against "V. bathopelagicus" Sal10, and 92.6% of LA51T against M. similis A3d10T, in maximum, respectively. Here, we show molecular phylogenetic, genomic, phenotypic, and chemotaxonomic features of the newly described species FE4T, FE10T, and LA51T. We also propose Pseudoalteromonas apostichopi sp. nov. with FE4T (JCM 36173 T = LMG 33143 T) as the type strain, Vibrio apostichopi sp. nov. with FE10T (JCM 36174 T = LMG 33144 T) as the type strain, and Marinobacter apostichopi sp. nov. with LA51T (JCM 36175 T = LMG 33145 T) as the type strain.
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Marinobacter , Filogenia , Pseudoalteromonas , Stichopus , Vibrio , Pseudoalteromonas/genética , Pseudoalteromonas/isolamento & purificação , Pseudoalteromonas/classificação , Animais , Vibrio/genética , Vibrio/classificação , Vibrio/isolamento & purificação , Stichopus/microbiologia , Marinobacter/genética , Marinobacter/classificação , Marinobacter/isolamento & purificação , Larva/microbiologia , Tipagem de Sequências Multilocus , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana , RNA Ribossômico 16S/genética , Zigoto/microbiologia , Genoma Bacteriano , Ácidos Graxos/análise , Ácidos Graxos/químicaRESUMO
BACKGROUND: Microbiome in early life has long-term effects on the host's immunological and physiological development and its disturbance is known to trigger various diseases in host Deuterostome animals. The sea cucumber Apostichopus japonicus is one of the most valuable marine Deuterostome invertebrates in Asia and a model animal in regeneration studies. To understand factors that impact on host development and holobiont maintenance, host-microbiome association has been actively studied in the last decade. However, we currently lack knowledge of early life core microbiome during its ontogenesis and how it benefits the host's growth. RESULTS: We analyzed the microbial community in 28 sea cucumber samples from a laboratory breeding system, designed to replicate aquaculture environments, across six developmental stages (fertilized eggs to the juvenile stage) over a three years-period to examine the microbiomes' dynamics and stability. Microbiome shifts occurred during sea cucumber larval ontogenesis in every case. Application of the most sophisticated core microbiome extraction methodology, a hybrid approach with abundance-occupancy core microbiome analyses (top 75% of total reads and > 70% occupation) and core index calculation, first revealed early life core microbiome consisted of Alteromonadaceae and Rhodobacteraceae, as well as a stage core microbiome consisting of pioneer core microbe Pseudoalteromonadaceae in A. japonicus, suggesting a stepwise establishment of microbiome related to ontogenesis and feeding behavior in A. japonicus. More interestingly, four ASVs affiliated to Alteromonadaceae and Rhodobacteraceae were extracted as early life core microbiome. One of the ASV (ASV0007) was affiliated to the Sulfitobactor strain BL28 (Rhodobacteraceae), isolated from blastula larvae in the 2019 raring batch. Unexpectedly, a bioassay revealed the BL28 strain retains a host growth-promoting ability. Further meta-pangenomics approach revealed the BL28 genome reads were abundant in the metagenomic sequence pool, in particular, in that of post-gut development in early life stages of A. japonicus. CONCLUSION: Repeated rearing efforts of A. japonicus using laboratory aquaculture replicating aquaculture environments and hybrid core microbiome extraction approach first revealed particular ASVs affiliated to Alteromonadaceae and Rhodobacteraceae as the A. japonicus early life core microbiome. Further bioassay revealed the growth promoting ability to the host sea cucumber in one of the core microbes, the Sulfitobactor strain BL28 identified as ASV0007. Genome reads of the BL28 were abundant in post-gut development of A. japonicus, which makes us consider effective probiotic uses of those core microbiome for sea cucumber resource production and conservation. The study also emphasizes the importance of the core microbiome in influencing early life stages in marine invertebrates. Understanding these dynamics could offer pathways to improve growth, immunity, and disease resistance in marine invertebrates.
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A Gram-staining-negative, oxidase-positive, strictly aerobic rod-shaped bacterium, designated strain PT1T, was isolated from the laboratory-reared larvae of the sea cucumber Apostichopus japonicus. A phylogenetic analysis based on the 16S rRNA gene nucleotide sequences revealed that PT1T was closely related to Neptuniibacter marinus ATR 1.1T (= CECT 8938T = DSM 100783T) and Neptuniibacter caesariensis MED92T (= CECT 7075T = CCUG 52065T) showing 98.2% and 98.1% sequence similarity, respectively. However, the average nucleotide identity (ANI) and in silico DNA-DNA hybridization (DDH) values among these three strains were 72.0%-74.8% and 18.3%-19.5% among related Neptuniibacter species, which were below 95% and 70%, respectively, confirming the novel status of PT1T. The average amino acid identity (AAI) values of PT1T showing 74-77% among those strains indicated PT1T is a new species in the genus Neptuniibacter. Based on the genome-based taxonomic approach, Neptuniibacter victor sp. nov. is proposed for PT1T. The type strain is PT1T (JCM 35563T = LMG 32868T).
Assuntos
Ácidos Graxos , Pepinos-do-Mar , Animais , Filogenia , Ácidos Graxos/análise , Larva/genética , RNA Ribossômico 16S/genética , Pepinos-do-Mar/genética , DNA , Análise de Sequência de DNA , DNA Bacteriano/genética , DNA Bacteriano/química , Técnicas de Tipagem Bacteriana , Hibridização de Ácido NucleicoRESUMO
The genus Thalassotalea is ubiquitous in marine environments, and up to 20 species have been described so far. A Gram-staining-negative, aerobic bacterium, designated strain PTE2T was isolated from laboratory-reared larvae of the Japanese sea cucumber Apostichopus japonicus. Phylogenetic analysis based on the 16S rRNA gene nucleotide sequences revealed that PTE2T was closely related to Thalassotalea sediminis N211T (= KCTC 42588T = MCCC 1H00116T) with 97.9% sequence similarity. ANI and in silico DDH values against Thalassotalea species were 68.5-77.0% and 19.7-24.6%, respectively, indicating the novelty of PTE2T. Based on genome-based taxonomic approaches, strain PTE2T (= JCM 34608T = KCTC 82592T) is proposed as a new species, Thalassotalea hakodatensis sp. nov.
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Ácidos Graxos , Pepinos-do-Mar , Animais , Filogenia , RNA Ribossômico 16S/genética , Pepinos-do-Mar/genética , Técnicas de Tipagem Bacteriana , Composição de Bases , Análise de Sequência de DNA , DNA Bacteriano/genética , Ubiquinona/genética , FosfolipídeosRESUMO
A Gram-staining-negative, aerobic bacterium, designated strain PT3T was isolated from laboratory-reared larvae of the Japanese sea cucumber Apostichopus japonicus. Phylogenetic analysis based on the 16S rRNA gene nucleotide sequences revealed that PT3T was closely related to Amphritea ceti RA1T (= KCTC 42154T = NBRC 110551T) and Amphritea spongicola MEBiC05461T (= KCCM 42943T = JCM 16668T) both with 98.3% sequence similarity, however, average nucleotide identity (ANI) and in silico DNA-DNA hybridization (in silico DDH) values among these three strains were below 95% and 70%, respectively, confirming the novelty of PT3T. Furthermore, the average amino acid identity (AAI) values of PT3T against other Amphritea species were on the reported genus delineation boundary (64-67%). Multilocus sequence analysis using four protein-coding genes (recA, mreB, rpoA, and topA) further demonstrated that PT3T, Amphritea ceti and Amphritea spongicola formed a monophyletic clade clearly separate from other members of the genus Amphritea. Three strains (PT3T, A. cetiãKCTC 42154T and A. spongicola JCM 16668T) also showed higher similarities in their core genomes compared to those of the other Amphritea spp. Based on the genome-based taxonomic approach, Aliamphritea gen. nov. was proposed together with the reclassification of the genus Amphritea and Aliamphritea ceti comb. nov. (type strain RA1T = KCTC 42154T = NBRC 110551T), Aliamphritea spongicola comb. nov. (type strain MEBiC05461T = KCCM 42943T = JCM 16668T), and Aliamphritea hakodatensis sp. nov. (type strain PT3T = JCM 34607T = KCTC 82591T) were suggested.
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Ácidos Graxos , Genômica , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Oceanospirillaceae , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
Sea cucumbers possess the remarkable capacity to regenerate their body parts or organs. Regeneration of host organs and/or body parts involves reconstruction of the host associated microbiota, however, the dynamics and contribution of microbiota to the regeneration process are largely unknown due to a lack of experimental models. To track the dynamics of individual gut microbiomes during gut regeneration, both caged mariculture and laboratory isolator systems of sea cucumbers (Apostichopus japonicus) were developed and longitudinal meta16S analyses were performed. Under natural environmental conditions in the caged mariculture system, both bacterial and eukaryotic communities in sea cucumbers' guts appeared to be reconstructed within 4 months after evisceration. Using the laboratory isolator, which can trace daily dynamics, we found that fecal microbiota collected before evisceration were clearly different from those collected after evisceration. We also identified eight key bacteria, belonging to Alteromonadaceae, Rhodobacteraceae, Oceanospirillaceae and family-unassigned Gammaproteobacteria, suggesting that these bacteria might interact with the host during the gut regeneration process. Six of the eight key bacteria were isolated for further bioassay using the isolator developed in this study to test whether these isolates affect gut regeneration.
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Mechanical stress stimulates bone remodeling, which occurs through bone formation and resorption, resulting in bone adaptation in response to the mechanical stress. Osteocytes perceive mechanical stress loaded to bones and promote bone remodeling through various cellular processes. Osteocyte apoptosis is considered a cellular process to induce bone resorption during mechanical stress-induced bone remodeling, but the underlying molecular mechanisms are not fully understood. Recent studies have demonstrated that neuropeptides play crucial roles in bone metabolism. The neuropeptide, methionine enkephalin (MENK) regulates apoptosis positively and negatively depending on cell type, but the role of MENK in osteocyte apoptosis, followed by bone resorption, in response to mechanical stress is still unknown. Here, we examined the roles and mechanisms of MENK in osteocyte apoptosis induced by compressive force. We loaded compressive force to mouse parietal bones, resulting in a reduction of MENK expression in osteocytes. A neutralizing connective tissue growth factor (CTGF) antibody inhibited the compressive force-induced reduction of MENK. An increase in osteocyte apoptosis in the compressive force-loaded parietal bones was inhibited by MENK administration. Nuclear translocation of NFATc1 in osteocytes in the parietal bones was enhanced by compressive force. INCA-6, which inhibits NFAT translocation into nuclei, suppressed the increase in osteocyte apoptosis in the compressive force-loaded parietal bones. NFATc1-overexpressing MLO-Y4 cells showed increased expression of apoptosis-related genes. MENK administration reduced the nuclear translocation of NFATc1 in osteocytes in the compressive force-loaded parietal bones. Moreover, MENK suppressed Ca2+ influx and calcineurin and calmodulin expression, which are known to induce the nuclear translocation of NFAT in MLO-Y4 cells. In summary, this study shows that osteocytes expressed MENK, whereas the MENK expression was suppressed by compressive force via CTGF signaling. MENK downregulated nuclear translocation of NFATc1 probably by suppressing Ca2+ signaling in osteocytes and consequently inhibiting compressive force-induced osteocyte apoptosis, followed by bone resorption. © 2020 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.
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Runt-related transcription factor 2 (Runx2) is an essential transcription factor for osteoblast differentiation, and is activated by mechanical stress to promote osteoblast function. Cleidocranial dysplasia (CCD) is caused by mutations of RUNX2, and CCD patients exhibit malocclusion and often need orthodontic treatment. However, treatment is difficult because of impaired tooth movement, the reason of which has not been clarified. We examined the amount of experimental tooth movement in Runx2+/- mice, the animal model of CCD, and investigated bone formation on the tension side of experimental tooth movement in vivo. Continuous stretch was conducted to bone marrow stromal cells (BMSCs) as an in vitro model of the tension side of tooth movement. Compared to wild-type littermates the Runx2+/- mice exhibited delayed experimental tooth movement, and osteoid formation and osteocalcin (OSC) mRNA expression were impaired in osteoblasts on the tension side of tooth movement. Runx2 heterozygous deficiency delayed stretch-induced increase of DNA content in BMSCs, and also delayed and reduced stretch-induced alkaline phosphatase (ALP) activity, OSC mRNA expression, and calcium content of BMSCs in osteogenic medium. Furthermore Runx2+/- mice exhibited delayed and suppressed expression of mammalian target of rapamycin (mTOR) and rapamycin-insensitive companion of mTOR (Rictor), essential factors of mTORC2, which is regulated by Runx2 to phosphorylate Akt to regulate cell proliferation and differentiation, in osteoblasts on the tension side of tooth movement in vivo and in vitro. Loss of half Runx2 gene dosage inhibited stretch-induced PI3K dependent mTORC2/Akt activity to promote BMSCs proliferation. Furthermore, Runx2+/- BMSCs in osteogenic medium exhibited delayed and suppressed stretch-induced expression of mTOR and Rictor. mTORC2 regulated stretch-elevated Runx2 and ALP mRNA expression in BMSCs in osteogenic medium. We conclude that Runx2+/- mice present a useful model of CCD patients for elucidation of the molecular mechanisms in bone remodeling during tooth movement, and that Runx2 plays a role in stretch-induced proliferation and osteogenesis in BMSCs via mTORC2 activation.
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We investigated an unknown ellipsoidal body that is sometimes found in the ovaries of the sea cucumber Apostichopus japonicus. Its external morphology, comprising an ellipsoidal dark central body (about 150 µm in length) and a surrounding transparent layer (about 50 µm in thickness), resembled that of a protozoan cyst, particularly an oocyst. Histological observations of the developing A. japonicus ovaries clarified that a small mass of organisms appeared in the cytoplasm of young oocytes, proliferated in these cells through budding, became rod shaped and arranged radially, and, finally, formed an outer layer. These processes were considered to be the formation of a cyst by a protozoan parasite. The small subunit ribosomal RNA (18S rRNA) gene was amplified from the DNA extracted from unknown ellipsoidal bodies by using polymerase chain reaction with universal primers for eukaryote 18S rRNA. The determined sequence was not identical to any of the known sequences in DNA databases, but it clustered in a clade of coccidian species belonging to Eucoccidiorida in phylogenetic analyses. From these results, we concluded that the unknown ellipsoidal body is a cyst (possibly an oocyst) of a coccidian parasite (order Eucoccidiorida) that is formed in the A. japonicus oocyte, though its lower taxonomic position is uncertain. In a survey of the gonads of wild A. japonicus at Esashi, Hokkaido, during the reproductive season, these cysts were detected in more than 50% of females but were never found in males. We consider that the cysts of this parasite can only be formed in A. japonicus ovaries.
Assuntos
Parasitos , Pepinos-do-Mar , Stichopus , Animais , Feminino , Masculino , Ovário , FilogeniaRESUMO
Deposit-feeding sea cucumbers repeat ingestion of sediments and excretion of faeces daily and consequently increase bacterial abundance in sediments and promote organic matter mineralization. Such ecological roles are expected to be collaborative activities of sea cucumbers and the gut microbiota. Here, we performed a spatiotemporally broad 16S rRNA gene analysis using 109 samples from sea cucumber faeces and habitat sediments to explore potential contribution of their gut microbiota to the ecological roles. Most operational taxonomic units (OTUs) observed in the faecal samples were shared with the sediment samples, nevertheless faecal and sediment microbiota differed from each other in UniFrac analysis. Lower bacterial diversity and increased relative abundance of specific OTUs in the faecal microbiota strongly suggest selective enrichment of ingested sediment microbiota in their guts. Interestingly, representative faecal OTUs were more abundant in sea cucumber-populated sediments than in un-inhabited sediments, indicating bacteria selectively enriched in the guts were spread on ambient sediments via faeces. Moreover, the predicted microbial community metabolic potential showed a higher abundance of genes related to carbohydrate and xenobiotics metabolisms in faeces than in sediments. Our study suggests the repeated selective enrichment transforms ambient sediment microbial communities and maintains the host's ecological roles by promoting organic matter mineralization.
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Microbioma Gastrointestinal , Trato Gastrointestinal/microbiologia , Pepinos-do-Mar/microbiologia , Animais , Análise por Conglomerados , DNA Ribossômico/química , DNA Ribossômico/genética , Fezes/microbiologia , Sedimentos Geológicos/microbiologia , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
Photoactive molecules with the frameworks of chlorin and/or porphyrin possessing four perfluorinated aromatic rings were conjugated with maltotriose (Mal3) via the nucleophilic aromatic substitution reaction and subsequent deprotection reaction of the oligosaccharide moieties. The resulting oligosaccharide-conjugated molecules are ultimately improved as compared to the previously reported monosaccharide-counterparts in terms of water-solubility. In particular, a water-soluble chlorin derivative surrounded by four Mal3 molecules showed an excellent biocompatibility, strong photoabsorption in the longer wavelength regions, and a very high photocytotoxicity. Thus, the present synthetic route combined with the use of an oligosaccharide was shown to be a straightforward strategy to develop a third generation photosensitizer for photodynamic therapy (PDT).
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Gut microbiome shapes various aspects of a host's physiology, but these functions in aquatic animal hosts have yet to be fully investigated. The sea cucumber Apostichopus japonicus Selenka is one such example. The large growth gap in their body size has delayed the development of intensive aquaculture, nevertheless the species is in urgent need of conservation. To understand possible contributions of the gut microbiome to its host's growth, individual fecal microbiome comparisons were performed. High-throughput 16S rRNA sequencing revealed significantly different microbiota in larger and smaller individuals; Rhodobacterales in particular was the most significantly abundant bacterial group in the larger specimens. Further shotgun metagenome of representative samples revealed a significant abundance of microbiome retaining polyhydroxybutyrate (PHB) metabolism genes in the largest individual. The PHB metabolism reads were potentially derived from Rhodobacterales. These results imply a possible link between microbial PHB producers and potential growth promotion in Deuterostomia marine invertebrates.
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Fezes/microbiologia , Hidroxibutiratos/metabolismo , Microbiota/genética , Stichopus/crescimento & desenvolvimento , Stichopus/microbiologia , Animais , Tamanho Corporal , Trato Gastrointestinal/microbiologia , Genes Bacterianos , MetagenomaRESUMO
This is a case report about the successful orthodontic treatment of a bilateral cleft lip and palate patient by using a combination of bone grafting and subsequent prosthodontic rehabilitation. An adult patient with a bilateral cleft lip and palate presented with a concave profile, anterior and lateral crossbite, a markedly deep overbite, and residual bilateral alveolar clefts. His jaw movement patterns were unstable and irregular due to his collapsed bite. Orthodontic treatment with bilateral bone grafting improved his concave profile by downward and backward rotation of the mandible within the freeway space, and optimum occlusion and functionally stable and smooth jaw movements were obtained. After a 6-year retention period, no skeletal relapse could be detected, and his occlusal stability was satisfactory.
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Transplante Ósseo , Fenda Labial/terapia , Fissura Palatina/terapia , Má Oclusão Classe II de Angle/terapia , Ortodontia Corretiva/métodos , Procedimentos de Cirurgia Plástica , Adulto , Anodontia/diagnóstico por imagem , Anodontia/terapia , Cefalometria , Fenda Labial/diagnóstico por imagem , Fissura Palatina/diagnóstico por imagem , Terapia Combinada , Humanos , Masculino , Má Oclusão Classe II de Angle/diagnóstico por imagem , Modelos Dentários , Radiografia PanorâmicaRESUMO
This study is an investigation of high-humidity aging effects on the total volatile organic compound (T-VOC) gas-sensing properties of platinum, palladium, and gold-loaded tin oxide (Pt,Pd,Au/SnO(2)) thick films. The sensor responses of the high-humidity aged Pt,Pd,Au/SnO(2), a non-aged Pt,Pd,Au/SnO(2), and a high-humidity aged Pt/SnO(2) to T-VOC test gas have been measured. The high-humidity aging is an effective treatment for resistance to humidity change for the Pt,Pd,Au/SnO(2) but not effective for the Pt/SnO(2). The mechanism of the high-humidity aging effects is discussed based on the change of surface state of the SnO(2) particles.
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Ouro/química , Umidade , Paládio/química , Platina/química , Compostos de Estanho/química , Compostos Orgânicos Voláteis/análiseRESUMO
Conventionally, the combination of mandibular and maxillary osteotomy is used to correct both mandibular deviation and maxillary canted occlusal plane. This case report describes functional and morphologic changes after an alternative treatment with miniscrew anchorage instead of maxillary osteotomy. A boy, aged 16 years 5 months, had temporomandibular joint disease symptoms, facial asymmetry with mandibular deviation, and a maxillary canted occlusal plane. The maxillary molars on the right side were extruded 3 mm compared with those on the left side. In addition, there were functional problems: lopsided chewing and asymmetric motion range of the bilateral condyles. After correction of the maxillary canted occlusal plane by intrusion of the maxillary right molars with miniscrew anchorage, the mandibular deviation was corrected with mandibular osteotomy. As the therapeutic result, functional symmetry was achieved in addition to symmetric morphologic changes, suggesting that combination treatment of miniscrew anchorage and mandibular osteotomy is effective for treating patients with mandibular deviation and maxillary canted occlusal plane.
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Assimetria Facial/cirurgia , Má Oclusão Classe III de Angle/terapia , Má Oclusão Classe II de Angle/terapia , Côndilo Mandibular/cirurgia , Procedimentos de Ancoragem Ortodôntica , Técnicas de Movimentação Dentária/métodos , Adolescente , Parafusos Ósseos , Cefalometria , Assimetria Facial/complicações , Humanos , Masculino , Má Oclusão Classe II de Angle/complicações , Má Oclusão Classe III de Angle/complicações , Maxila , Ortodontia Corretiva/instrumentação , Ortodontia Corretiva/métodos , Osteotomia , Resultado do TratamentoRESUMO
While it has been reported that the blood level of dioxins and polychlorinated biphenyls in humans may be decreased by treatment with colestimide, the effects of the agent are still obscure. To address this issue, we examined the effect of Cholebine, a cholesterol lowering agent containing colestimide as an active ingredient, on the excretion of 2,3,4,7,8-pentachlorodibenzofuran (PenCDF) in rats. In a short term study, male Wistar rats (5 weeks-old) were given chows including 3% Cholebine (PenCDF/Chol group) or control chows (PenCDF group) for 7 days after administration of 14C-labeled PenCDF (0.5 mg/kg body weight, p.o.). On day 1, the fecal excretion of PenCDF in the PenCDF/Chol group was greater by 15% compared to that of the PenCDF group. Although some increases were also observed during day 2 and day 7, Cholebine did not exhibit any marked effect on the fecal excretion of PenCDF. The tissue concentrations of PenCDF at day 7 in the PenCDF/Chol group showed a 20%-30% decrease compared with those of the PenCDF group, except that the level in the brain was comparable between the two groups. The fecal excretion of PenCDF in a long term study (Cholebine treatment for 28 days) demonstrated the same tendency as that of short-term study. However, in long-term study, the Cholebine had no effect on the tissue concentration of PenCDF except for brain. An increase in PenCDF excretion by Cholebine seems to be due to the binding of Cholebine to bile acids as lipid carriers. This is because no binding of Cholebine to 14C-PenCDF was detected. These results suggest that Cholebine has little effect on the reabsorption of dioxin, whereas it reduces substantially the first absorption of dioxin.
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Anticolesterolemiantes/farmacologia , Benzofuranos/metabolismo , Ácidos e Sais Biliares/farmacologia , Dioxinas/metabolismo , Animais , Benzofuranos/análise , Fezes/química , Masculino , Ratos , Ratos WistarRESUMO
A female patient with a skeletal Class III severe anterior openbite was treated using miniplates as the anchorage. The patient was 15 years and 10 months of age when she reported to our university hospital with a chief complaint of anterior openbite and reversed occlusion. The patient had an anterior openbite with an overjet of -3.0 mm and overbite of -5.0 mm and a Class III molar relationship. The cephalometric analysis showed a skeletal Class III relationship (ANB 0 degrees ). After the extraction of the bilateral mandibular third molars, miniplates were placed in the mandibular external oblique line. The mandibular dentition was retracted using elastic chain and miniplates. After treatment, an Angle Class I molar relationship was achieved and overjet and overbite had become 2.0 mm and 1.5 mm. A good facial appearance and occlusal relationship were obtained. The total active orthodontic treatment period was 23 months. Wrap-around type retainers were placed on both jaws and a lingual bonded retainer was also attached in the mandibular incisors. After 1 year of retention, the occlusion was stable, and a good facial profile was also retained. The mandibular deviation to the left was improved and the strain in the circumoral musculature during lip closure disappeared. An appropriate interincisal relationship was achieved by the uprighting of mandibular dentition without changing the vertical intermaxillary relationship. A panoramic radiograph showed no marked root resorption. Our results suggest that implant anchorage is useful for correction of skeletal Class III severe anterior openbite cases.
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Má Oclusão Classe III de Angle/terapia , Mordida Aberta/terapia , Procedimentos de Ancoragem Ortodôntica/métodos , Adolescente , Placas Ósseas , Cefalometria , Arco Dental/patologia , Estética Dentária , Feminino , Humanos , Má Oclusão Classe III de Angle/patologia , Dente Serotino/cirurgia , Mordida Aberta/patologia , Procedimentos de Ancoragem Ortodôntica/instrumentação , Contenções Ortodônticas , Técnica de Expansão Palatina , Extração Seriada , Técnicas de Movimentação Dentária/instrumentação , Técnicas de Movimentação Dentária/métodos , Resultado do TratamentoRESUMO
INTRODUCTION: Skeletal anterior open bite is a difficult problem to correct in orthodontic treatment. In adults, treatment of severe skeletal anterior open bite consists mainly of surgically repositioning the maxilla or the mandible. Recently, molar intrusion by using skeletal anchorage has been developed as a new strategy for open-bite treatment. In this study, we compared treatment outcomes in patients with severe anterior open bite treated with molar intrusion by using skeletal anchorage and with orthognathic surgery. METHODS: Twenty-three subjects with overbite less than -3.0 mm were treated with skeletal anchorage (n = 10) or with LeFort I osteotomy combined with mandibular osteotomy (n = 13). Pretreatment and posttreatment lateral cephalograms were compared. RESULTS: Incisors were significantly elongated in the surgically treated subjects (4.6 mm, P <.01). There were no significant differences in the treatment results between skeletal anchorage and surgery, with reduced facial heights of 4.0 and 3.8 mm, and increased overbites of 6.8 and 7.0 mm, respectively. CONCLUSIONS: These results suggest that molar intrusion with skeletal anchorage is simpler and more useful than 2-jaw surgery in the treatment of patients with severe anterior open bite.