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Abstract LeuO plays the role of a master regulator in the cyclic-L-phenylalanine-L-proline (cFP)-dependent signaling pathway in Vibrio vulnificus. cFP, as shown through isothermal titration calorimetry analysis, binds specifically to the periplasmic domain of ToxR. Binding of cFP triggers a change in the cytoplasmic domain of ToxR, which then activates transcription of leuO encoding a LysR-type regulator. LeuO binds to the region upstream of its own coding sequence, inhibiting its own transcription and maintaining a controlled level of expression. A five-bp deletion in this region abolished expression of LeuO, but a ten-bp deletion did not, suggesting that a DNA bending mechanism is involved in the regulation. Furthermore, binding of RNA polymerase was significantly lower both in the deletion of the ToxR binding site and in the five-bp deletion, but not in the ten-bp deletion, as shown in pull-down assays using an antibody against RNA polymerase subunit α. In summary, multiple factors are involved in control of the expression of LeuO, a master regulator that orchestrates downstream regulators to modulate factors required for survival and pathogenicity of the pathogen.

Src-homology-2-containing phosphotyrosine phosphatase (SHP2), a classic cytoplasmic protein and a major regulator of receptor tyrosine kinases and G protein-coupled receptors, plays a significant role in preimplantation embryo development. In this study, we deciphered the role of SHP2 in the somatic compartment of oocytes during meiotic maturation. SHP2 showed nuclear/cytoplasmic localization in bovine cumulus and human granulosa (COV434) cells. Follicle-stimulating hormone (FSH) treatment significantly enhanced cytoplasmic SHP2 localization, in contrast to the E2 treatment, which augmented nuclear localization. Enhanced cytoplasmic SHP2 was found to negatively regulate the expression of the ERα-transcribed NPPC and NPR2 mRNAs, which are vital for oocyte meiotic arrest. The co-immunoprecipitation results revealed the presence of the SHP2/ERα complex in the germinal vesicle-stage cumulus–oocyte complexes, and this complex significantly decreased with the progression of meiotic maturation. The complex formation between ERα and SHP2 was also confirmed by using a series of computational modeling methods. To verify the correlation between SHP2 and NPPC/NPR2, SHP2 was knocked down via RNA interference, and NPPC and NPR2 mRNAs were analyzed in the control, E2, and FSH-stimulated COV434 cells. Furthermore, phenyl hydrazonopyrazolone sulfonate 1, a site-directed inhibitor of active SHP2, showed no significant effect on the ERα-transcribed NPPC and NPR2 mRNAs. Taken together, these findings support a novel nuclear/cytoplasmic role of SHP2 in oocyte meiotic resumption and maturation.

Immune adaptation plays an essential role in determining pregnancy, which has been shown to be dependent on sufficient immunological tolerance mediated by FOXP3+ regulatory T cells. Recently, an X-linked maternal single-nucleotide polymorphism (SNP), located 2175 base pairs upstream of the start codon in the bovine FOXP3 gene (NC_037357.1: g.87298881A>G, rs135720414), was identified in Japanese Black (JB: Bos taurus) cows in association with recurrent infertility. However, with the exception of JB cows, the frequency of this SNP has yet to be studied in other cow populations. In this study, we thus aimed to evaluate the frequency of this SNP in different cow breeds. Between 2018 and 2021, a total of 809 DNA samples were obtained from 581 JB, 73 Holstein Friesian (HF: B. taurus), 125 Korean Hanwoo (KH: B. taurus coreanae), and 30 Indonesian Madura (IM: a crossbreed between B. indicus and B. javanicus) cows, which were genotyped using a TaqMan probe-based real-time polymerase chain reaction assay designed in this study. The frequency of the G allele was found to be relatively high in local IM (0.700), moderate in dairy HF (0.466), and low in beef JB (0.250) and KH (0.112) cows, with differences in the frequencies between each group being shown to be statistically significant (p < 0.005) using Fisher’s exact test. The results obtained in this study indicate that the G allele frequencies of the identified the SNP differ markedly in different breeds of taurine and indicine cattle. Given these findings, it would thus be important to evaluate the relationships between high frequencies of the G allele and infertility in different breeds.

Normal adult tissue fibroblasts can be activated during wound healing, pathologic fibrosis and in cancer stroma, but the regulatory network that controls its dynamics is unknown. Here the authors show that fibroblasts are activated by a positive feedback loop formed by Twist1, Prrx1, and Tenascin-C bi-stably.

Zirconia implants have become an alternative to titanium implants due to several advantages. The zirconia implant is relatively esthetic and highly resistant to bacteria. While biomaterial studies for zirconia implants have considerably accumulated, in vivo studies have not yet progressed. In the present study, the functional and biological properties of zirconia implants were analyzed thorough in vitro and in vivo studies. The proliferation properties of periodontal cells on the discs of machined surface titanium, hydroxyapatite coated titanium and zirconia were analyzed, and zirconia was shown to be favorable. In addition, small implant fixtures that can be applied to the jawbone of mice were manufactured and transplanted to C57BL/6 mice. The adhesion molecules expression patterns in peri-implant mucosa suggest a stronger mucosal seal and more adequate prevention of peri-implant epithelium (PIE) elongation in the zirconia implant when compared with other conventional materials. Differential laminin-332 expression in peri-implant mucosa of zirconia implants seems to regulate the PIE elongation. In conclusion, zirconia was found to be promising and advantageous with regards to the mucosal seal. And biological width (BW) of peri-implant mucosa is more desirable in zirconia implants compared to conventional titanium implants.

Reproduction of the exact interocclusal relationship using digital workflow is crucial for precise fabrication of accurate prostheses. Intraoral scanner is known to be valid for the measurement of quadrants, however, the role of missing area in the quadrant scan on the virtual interocclusal record (VIR) is uncertain. This study aimed to evaluate the accuracy of VIR in quadrant scans using an intraoral scanner (IOS) under four different edentulous conditions. Eight scans per group were obtained using a laboratory scanner and three IOSs (Trios3, CS3600, i500). Based on trueness and precision, Trios3 had the best results, followed by CS3600 and i500. The trueness and precision were affected by edentulous conditions. The three IOSs showed deviation in the posterior region during assessment of VIR for the missing area with posterior support. CS3600 and i500 showed deviation in the short-span edentulous area without support. In extended edentulous condition without support, Trios3 showed overclosure, while i500 showed an angular deviation. In some groups scanned with Trios3 and i500, the tilting effect was observed. Based on the edentulous condition and type of IOS used, local or general deviations in occlusion were seen. The accuracy of VIR was dependent on accurate scan data. Thus, registration of the occlusal relationship in an edentulous area with more than two missing teeth using IOSs may be clinically more inaccurate than that with a laboratory scanner.

Brassica oleracea var. capitata L. (cabbage) is a popular vegetable with a wide range of pharmacological activities that help to promote human health. The present study investigated the beneficial effects of B. oleracea var. capitata L. extract (BOE) on HCl/ethanol (H/E)-induced gastric damages in mice. Pre-administration of BOE (25–100 mg/kg) for 7 consecutive days significantly decreased macroscopically visible lesion on the gastric mucosa induced by H/E. In addition, results from hematoxylin and eosin-stained gastric tissue showed that BOE inhibited invaded percentage of lesion and prevented the reduction in mucosal thickness in peri-ulcerative region. BOE significantly alleviated the H/E-mediated decreases in Alcian blue binding, total hexose, sialic acid, and collagen in the gastric tissue, suggesting BOE attenuates the gastric damage via preserving the integrity of gastric mucus. Moreover, BOE significantly decreased histamine level in the plasma and reduced mRNA levels associated with secreting gastric acid. Furthermore, BOE inhibited myeloperoxidase activity and suppressed nuclear factor-κB mRNA and its dependent inflammatory genes expression induced by H/E. BOE also strengthened antioxidant enzyme activity, with a mitigating H/E-mediated increase in malondialdehyde level of the gastric tissue. Thus, these results suggest that BOE has the potential to protect the gastric tissue via inhibiting gastric acid secretion, inflammation, and oxidative stress.

Recently, digital technology has been used in dentistry to enhance accuracy and to reduce operative time. Due to advances in digital technology, the integration of individual mandibular motion into the mapping of the occlusal surface is being attempted. The Patient Specific Motion (PSM) is one such method. However, it is not clear whether the occlusal design that is adjusted using PSM could clinically show reduced occlusal error compared to conventional methods based on static occlusion. In this clinical comparative study including fifteen patients with a single posterior zirconia crown treatment, the occlusal surface after a clinical adjustment was compared to no adjustment (NA; design based on static occlusion), PSM (adjusted using PSM), and adjustment using a semi-adjustable articulator (SA) for the assessment of occlusal error. The root mean square (RMS; μm), average deviation value (±AVG; μm), and proportion inside the tolerance (in Tol; %) were calculated using the entire, subdivided occlusal surface and the out of tolerance area. Using a one-way ANOVA, the RMS and +AVG from the out of tolerance area showed a statistical difference between PSM (202.3 ± 39.8 for RMS, 173.1 ± 31.3 for +AVG) and NA (257.0 ± 73.9 for RMS, 210.9 ± 48.6 for +AVG). For the entire and subdivided occlusal surfaces, there were no significant differences. In the color-coded map analysis, PSM demonstrated a reduced occlusal error compared to NA. In conclusion, adjustment occlusal design using PSM is a simple and effective method for reducing occlusal errors that are difficult to identify in a current computer-aided design (CAD) workflow with static occlusion.

Recent advances in three-dimensional (3D) printing have introduced new materials that can be utilized for dental restorations. Nonetheless, there are limited studies on the color stability of restorations using 3D-printed crowns and bridge resins. Herein, the color stability of conventional computer-aided design/computer-aided manufacturing (CAD/CAM) blocks and 3D-printing resins was evaluated and assessed for their degrees of discoloration based on material type, colorant types (grape juice, coffee, curry, and distilled water (control group)), and storage duration (2, 7, and 30 days) in the colorants. Water sorption, solubility, and scanning electron microscope (SEM) analyses were conducted. A three-way ANOVA analysis showed that all three factors significantly affected the color change of the materials. Notably, the discoloration (ΔE00) was significantly higher in all 3D printing resins (4.74–22.85 over the 30 days) than in CAD/CAM blocks (0.64–4.12 over the 30 days) following immersion in all colorants. 3D-printing resins showed color differences above the clinical limit (2.25) following storage for 7 days or longer in all experimental groups. Curry was the most prominent colorant, and discoloration increased in almost all groups as the storage duration increased. This study suggests that discoloration must be considered when using 3D printing resins for restorations.

Studies on the constructability of cold-formed steel (CFS) members are lacking. In this context, this paper proposes an integrated panel to replace the studs and steel sheet in the steel-sheathed wall system used in light gauge steel frames. The integrated panel was developed, to not only exhibit better structural performance and constructability than the steel-sheathed wall system, but to also reflect the appropriate details for applications to on-site panel construction. Cyclic loading tests were performed to investigate the seismic performances of three integrated panel specimens and a steel-sheathed panel specimen. The integrated panel specimens exhibited greater deformation capacity, ductility and equivalent damping ratio than the steel-sheathed panel. Although the web slenderness of the integrated member was nearly 400, the nominal strength of the integrated panel was predictable on an AISI standard. However, for stud-reinforced integrated panels, the nominal strength was overestimated by 45%−60%, and a draft equation was proposed to predict the yield strength of these panels. The proposed equation accurately predicted the yield strength of the stud-reinforced integrated panel specimens. The study results can aid in the construction of light gauge steel frames that require reinforcing parts with a high web slenderness ratio.

We describe a novel insulin-degrading enzyme, SidC, that contributes to the proliferation of the human bacterial pathogen Vibrio vulnificus in a mouse model. SidC is phylogenetically distinct from other known insulin-degrading enzymes and is expressed and secreted specifically during host infection. Purified SidC causes a significant decrease in serum insulin levels and an increase in blood glucose levels in mice. A comparison of mice infected with wild type V. vulnificus or an isogenic sidC deletion strain showed that wild type bacteria proliferated to higher levels. Additionally, hyperglycemia leads to increased proliferation of V. vulnificus in diabetic mice. Consistent with these observations, the sid operon was up-regulated in response to low glucose levels through binding of the cAMP-receptor protein (CRP) complex to a region upstream of the operon. We conclude that glucose levels are important for the survival of V. vulnificus in the host, and that this pathogen uses SidC to actively manipulate host endocrine signals, making the host environment more favorable for bacterial survival and growth. [ABSTRACT FROM AUTHOR]