Study on the virulence factors of Vibrio vulnificus by transposon insertion mutagenesis

2015岡山県立大学大学

Lip incompetence and myofunctional therapy

Lip incompetence, which is a state in which the upper and lower lips are constantly apart, is thought to induce mouth breathing that may result in the development of gingivitis and periodontal disease. In the orthodontic field, lip incompetence has major problems for diagnosis, when determining the treatment goal and at the time of retention. We have been performing studies on lip incompetence and have obtained the following results : (1) it is possible to predict to some extent the state of lip closure during sleep by examination only in an awake state, (2) anterior teeth are positioned forward in individuals with lip incompetence, and (3) a training method called Button Pull is effective as a myofunctional therapy for improving lip incompetence, but there remains a need to clarify the background

A deazariboflavin chromophore kinetically stabilizes reduced FAD state in a bifunctional cryptochrome

An animal-like cryptochrome derived from Chlamydomonas reinhardtii (CraCRY) is a bifunctional flavoenzyme harboring flavin adenine dinucleotide (FAD) as a photoreceptive/catalytic center and functions both in the regulation of gene transcription and the repair of UV-induced DNA lesions in a light-dependent manner, using different FAD redox states. To address how CraCRY stabilizes the physiologically relevant redox state of FAD, we investigated the thermodynamic and kinetic stability of the two-electron reduced anionic FAD state (FADH−) in CraCRY and related (6–4) photolyases. The thermodynamic stability of FADH− remained almost the same compared to that of all tested proteins. However, the kinetic stability of FADH− varied remarkably depending on the local structure of the secondary pocket, where an auxiliary chromophore, 8-hydroxy-7,8-didemethyl-5-deazariboflavin (8-HDF), can be accommodated. The observed effect of 8-HDF uptake on the enhancement of the kinetic stability of FADH− suggests an essential role of 8-HDF in the bifunctionality of CraCRY.Hosokawa Y., Morita H., Nakamura M., et al. A deazariboflavin chromophore kinetically stabilizes reduced FAD state in a bifunctional cryptochrome. Scientific Reports 13, 16682 (2023); https://doi.org/10.1038/s41598-023-43930-0

Comparison of Protein Profiles of Gingival Crevicular Fluids Collected from Incisors, Canines, and Molars

Many studies have shown that gingival crevicular fluid （GCF） reflects the inflammatory state of local periodontal tissues. GCF has been collected from several types of teeth in previous studies. However, there is no report that characterizes GCF by the type of tooth. In the present study, the protein profiles of GCF from different sites were comprehensively compared with each other. GCF was sampled from six healthy adult men （21-31 years old） with healthy periodontal tissues. Three separate GCF samples were collected at the maxillary central incisor, canine, and first molar of each individual. The protein profiles of GCF were analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis and liquid chromatogram-tandem mass spectrometry （LC-MS/MS）. The band patterns on the sodium dodecyl sulfate polyacrylamide gel electrophoresis from the set of three GCF samples from each individual were similar, regardless of the type of tooth. The proteins contained in each band were identified by LC-MS/MS analysis, and they were found to be the same among the three GCF samples. A comprehensive and quantitative analysis of proteins in the GCF samples was performed by LC-MS/MS using isobaric tag labeling. In total, 86 proteins were identified in GCF. A small number of proteins were increased or decreased in GCF from the first molars compared with the other types of teeth in one or two individuals. However, overall, no proteins were found to exhibit a reproducibly different composition in any of the individuals. These analyses show that the protein profiles of GCF in healthy periodontal tissues are similar, regardless of the type of tooth

Effect of Sensory Manipulations on Human Joint Stiffness Strategy and Its Adaptation for Human Dynamic Stability

Sensory input plays an important role to human posture control system to initiate strategy in order to counterpart any unbalance condition and thus, prevent fall. In previous study, joint stiffness was observed able to describe certain issues regarding to movement performance. But, correlation between balance ability and joint stiffness is still remains unknown. In this study, joint stiffening strategy at ankle and hip were observed under different sensory manipulations and its correlation with conventional clinical test (Functional Reach Test) for balance ability was investigated. In order to create unstable condition, two different surface perturbations (tilt up-tilt (TT) down and forward-backward (FB)) at four different frequencies (0.2, 0.4, 0.6 and 0.8 Hz) were introduced. Furthermore, four different sensory manipulation conditions (include vision and vestibular system) were applied to the subject and they were asked to maintain their position as possible. The results suggested that joint stiffness were high during difficult balance situation. Less balance people generated high average joint stiffness compared to balance people. Besides, adaptation of posture control system under repetitive external perturbation also suggested less during sensory limited condition. Overall, analysis of joint stiffening response possible to predict unbalance situation faced by human

Analysis of Clinical Outcome of Patients with Poorly Differentiated Thyroid Carcinoma

Background. We retrospectively analyzed whether poor differentiation is the independent prognostic factor for thyroid carcinoma or not. Methods. The subjects were 29 patients with PDTC who were treated between April 1996 and March 2006 to compare with those of well-differentiated papillary carcinoma patients (n = 227). Results. The relapse free (RFS), distant relapse-free survival and cause-specific survival, rates were significantly lower in patients with PDTC (P < .0001, P < .001, and P < .05). After classification into focal (<10%) and diffuse type (over 10%) of PDTC, there were no significant differences in RFS and cause-specific survival due to component type or proportion of poorly differentiated component. On multivariate analysis, poor differentiation (P < .0005, RR = 4.456, 95% CI; 1.953–10.167) and extrathyroidal infiltration (P < .05, RR = 2.898, 95% CI; 1.278–6.572) showed a significant impact on DFS, and poor differentiation (P < .05, RR = 9.343, 1.314–66.453) and age (P < .005, RR = 1.306, 1.103–1.547) significantly impacted cause-specific survival. Conclusion. Poor differentiation was an independent factor for survival. Distant relapse was significantly more common among PDTC patients, and systemic therapy might be warranted

Tissue-Tissue Interaction-Triggered Calcium Elevation Is Required for Cell Polarization during Xenopus Gastrulation

The establishment of cell polarity is crucial for embryonic cells to acquire their proper morphologies and functions, because cell alignment and intracellular events are coordinated in tissues during embryogenesis according to the cell polarity. Although much is known about the molecules involved in cell polarization, the direct trigger of the process remains largely obscure. We previously demonstrated that the tissue boundary between the chordamesoderm and lateral mesoderm of Xenopus laevis is important for chordamesodermal cell polarity. Here, we examined the intracellular calcium dynamics during boundary formation between two different tissues. In a combination culture of nodal-induced chordamesodermal explants and a heterogeneous tissue, such as ectoderm or lateral mesoderm, the chordamesodermal cells near the boundary frequently displayed intracellular calcium elevation; this frequency was significantly less when homogeneous explants were used. Inhibition of the intracellular calcium elevation blocked cell polarization in the chordamesodermal explants. We also observed frequent calcium waves near the boundary of the dorsal marginal zone (DMZ) dissected from an early gastrula-stage embryo. Optical sectioning revealed that where heterogeneous explants touched, the chordamesodermal surface formed a wedge with the narrow end tucked under the heterogeneous explant. No such configuration was seen between homogeneous explants. When physical force was exerted against a chordamesodermal explant with a glass needle at an angle similar to that created in the explant, or migrating chordamesodermal cells crawled beneath a silicone block, intracellular calcium elevation was frequent and cell polarization was induced. Finally, we demonstrated that a purinergic receptor, which is implicated in mechano-sensing, is required for such frequent calcium elevation in chordamesoderm and for cell polarization. This study raises the possibility that tissue-tissue interaction generates mechanical forces through cell-cell contact that initiates coordinated cell polarization through a transient increase in intracellular calcium

The Expression Level of HIV-1 Vif Is Optimized by Nucleotide Changes in the Genomic SA1D2prox Region during the Viral Adaptation Process

HIV-1 Vif plays an essential role in viral replication by antagonizing anti-viral cellular restriction factors, a family of APOBEC3 proteins. We have previously shown that naturally-occurring single-nucleotide mutations in the SA1D2prox region, which surrounds the splicing acceptor 1 and splicing donor 2 sites of the HIV-1 genome, dramatically alter the Vif expression level, resulting in variants with low or excessive Vif expression. In this study, we investigated how these HIV-1 variants with poor replication ability adapt and evolve under the pressure of APOBEC3 proteins. Adapted clones obtained through adaptation experiments exhibited an altered replication ability and Vif expression level compared to each parental clone. While various mutations were present throughout the viral genome, all replication-competent adapted clones with altered Vif expression levels were found to bear them within SA1D2prox, without exception. Indeed, the mutations identified within SA1D2prox were responsible for changes in the Vif expression levels and altered the splicing pattern. Moreover, for samples collected from HIV-1-infected patients, we showed that the nucleotide sequences of SA1D2prox can be chronologically changed and concomitantly affect the Vif expression levels. Taken together, these results demonstrated the importance of the SA1D2prox nucleotide sequence for modulating the Vif expression level during HIV-1 replication and adaptation