Inhibition of ATR Increases the Sensitivity to WEE1 Inhibitor in Biliary Tract Cancer

Purpose Currently, the DNA damage response (DDR) pathway represents a key target for new cancer drug development. Advanced biliary tract cancer (BTC) has a poor prognosis because of the lack of efficacious treatment options. Although DNA repair pathway alterations have been reported in many patients with BTC, little is known regarding the effects of DDR-targeted agents against BTC. Materials and Methods In this study, nine BTC cell lines were exposed to the WEE1 inhibitor (AZD1775). In vitro, MTT assay, colony-forming assay, cell cycle analysis, phospho-histone H3 staining assay, Transwell migration assay, and western blot were performed. Then, to enhance the antitumor effect of AZD1775, the combination treatment of WEE1 inhibitor and ataxia telangiectasia mutated and Rad3 related (ATR) inhibitor (AZD6738) was conducted using MTT assay and comet assay. Finally, HuCCT-1 and SNU2670 xenograft models were established to confirm the anti-tumor effect of AZD1775 alone. Furthermore, the combination treatment was also evaluated in SNU2670 xenograft models. Results AZD1775 blocked the phosphorylation of CDC2 and CDC25C in all cell lines, but significantly increased apoptosis and S phase arrest in sensitive cells. However, increased p-ATR and phosphorylated ataxia telangiectasia mutated levels were observed in less sensitive cells. In addition, in vitro and in vivo data illustrated that AZD1775 combined with AZD6738 exerted more potent anti-tumor effects than either drug alone. Although WEE1 inhibition has promising anti-tumor effects in some BTC cells, the addition of ATR inhibitors could enhance its efficacy. Conclusion Taken together, this study supports further clinical development of DDR-targeted strategies as monotherapy or combination regimens for BTC.

Contribution of actin filaments and microtubules to cell elongation and alignment depends on the grating depth of microgratings

Additional file 1: Figure S1. (A) A phase contrast image of TCPS surface. Bar, 100 μm. (B) An imageshowing FN-lines (1 μm line and spacing) obtained by Atomic Force Microscopy (AFM) (Dimension 3100with a Nanoscope III controller, Digital Instruments) using silicon cantilevers (spring constant; 50 Nm-1)(RTESP, Veeco Probes) in contact mode. (C-E) SEM (Scanning electron microscopy) (6010 LV, JEOL)images showing the cross section of three different microgratings; 1 μm gratings with 0.35 um depth (C) and1 μm depth (D) and 2 μm gratings with 2 μm depth (E). Figure S2. (A) Fluorescence image of a RPE-1 cell stably expressing GFP/centrin cell on 1 μm gratings (1 μm deep). Bar, 30 μm. A yellow arrow indicates the direction of cell elongation. (B) Average cell aspect ratio (R) of cells on 1 μm gratings (0.35 or 1 μm deep) and 2 μm gratings with/without CD treatment. n: number of cells. ***P < 0.001. Data were analyzed using one-way ANOVA and a Bonferroni post hoc test. Error bar denotes the standard deviation of the mean. Figure S3. Alignment of actin and vinculin to the different substrates (Flat TCPS surface, FN-lines, and 1 μm gratings (0.35 or 1μm deep)). The alignment angle was measured as an angle difference of actin or vinculin orientation to the long axis of a cell on flat PDMS surface or the long axis of the FN-line or each micrograting. #: the number of cells. Error bar denotes the standard deviation of the mean. Figure S4. Merged image of MTs (Green fluorescence) and pattern (phase contrast) of cells on 1 μm grating (1 μm deep) in the presenceof CD at 1 μM

Changes in the bony alignment of the foot after tendo-Achilles lengthening in patients with planovalgus deformity

Background This study was performed to investigate the change in the bony alignment of the foot after tendo-Achilles lengthening (TAL) and the factors that affect these changes in patients with planovalgus foot deformity. Methods Consecutive 97 patients (150 feet; mean age 10 years; range 5.1–35.7) with Achilles tendon contracture (ATC) and planovalgus foot deformity who underwent TAL were included. All patients underwent preoperative and postoperative weight-bearing anteroposterior (AP) or lateral (LAT) foot radiographics. Changes in AP talo-1st metatarsal angle, AP talo-2nd metatarsal angle, LAT talo-1st metatarsal angle, and calcaneal pitch angle and the factors affecting such changes after TAL were analyzed using lineal mixed model. Results There were no significant change in AP talo-1st metatarsal angle and AP talo-2nd metatarsal angle after TAL in patients with cerebral palsy (CP) (p = 0.236 and 0.212). However, LAT talo-1st metatarsal angle and calcaneal pitch angle were significantly improved after TAL (13.0°, p < 0.001 and 4.5°, p < 0.001). Age was significantly associated with the change in LAT talo-1st metatarsal angle after TAL (p = 0.028). The changes in AP talo-1st metatarsal angle, AP talo-2nd metatarsal angle, and calcaneal pitch angle after TAL were not significantly associated with the diagnosis (p = 0.879, 0.903, and 0.056). However, patients with CP showed more improvement in LAT talo-1st metatarsal angle (− 5.0°, p = 0.034) than those with idiopathic cause. Conclusion This study showed that TAL can improve the bony alignment of the foot in patients with planovalgus and ATC. We recommend that physicians should consider this studys findings when planning operative treatment for such patients.This study was supported by the National Research Foundation of Korea(NRF) grant funded by the Korea government (MSIT) (No. NRF2019R1C1C1010352)

Identification of marbling-related candidate genes in M-longissimus dorsi of high- and low marbled Hanwoo (Korean native cattle) steers

This study was conducted to identify marbling-related candidate genes in M. longissimus dorsi of high- and low-marbled Hanwoo. The longissimus dorsi muscles were selected for gene expression from eight Hanwoo steer carcasses based on crude fat content. In the analysis of variance, gene expression of five candidate genes, FABP4, SCD, PPARγ, Titin and Nebulin was determined to be significantly different between high- and low-marbled Hanwoo steers (P < 0.0001). The Pik-4 and CaMK II genes were also shown to have a significant effect on crude fat content (P < 0.01). In the analysis of the differential expression between high- and low marbled groups, FABP4 gene expression was approximately 2 times higher in the high marbled group relative to the low marbled group. However, the PPAR y and SCD gene were highly expressed in the low marbled group. In addition, Titin and Nebulin were highly expressed in the low marbled group when placed under relatively high shear force. Finally, the Pik-4 and CaMKII gene also displayed a high expression pattern in the low marbled grou

Effect of bioactive glass addition on the physical properties of mineral trioxide aggregate

Background The addition of bioactive glass (BG), a highly bioactive material with remineralization potential, might improve the drawback of weakening property of mineral trioxide aggregates (MTA) when it encounters with body fluid. This study aims to evaluate the effect of BG addition on physical properties of MTA. Methods ProRoot (MTA), and MTA with various concentrations of BG (1, 2, 5 and 10% BG/MTA) were prepared. Simulated body fluid (SBF) was used to investigate the effect of the storage solution on dentin remineralization. Prepared specimens were examined as following; the push-out bond strength to dentin, compressive strength, setting time solubility and X-ray diffraction (XRD) analysis. Results The 2% BG/MTA showed higher push-out bond strengths than control group after 7 days of SBF storage. The 2% BG/MTA exhibited the highest compressive strength. Setting times were reduced in the 1 and 2% BG/MTA groups, and solubility of all experimental groups were clinically acceptable. In all groups, precipitates were observed in dentinal tubules via SEM. XRD showed the increased hydroxyapatite peaks in the 2, 5 and 10% BG/MTA groups. Conclusion It was verified that the BG-added MTA increased dentin push-out bond strength and compressive strength under SBF storage. The addition of BG did not negatively affect the MTA maturation reaction; it increased the amount of hydroxyapatite during SBF maturation.This research was supported by the Bio & Medical Technology Development Program of the National Research Foundation (NRF) & funded by the Korean government (MSIP&MOHW) (No. 2017M3A9E4048170) and NRF (No.2019R1G1A1100082). This work was also supported by a grant from Kyung Hee University in 2020 (KHU-20201233)