Journal: PLoS ONE

Article Title: Periodontal status and the incidence of selected bacterial pathogens in periodontal pockets and vascular walls in patients with atherosclerosis and abdominal aortic aneurysms

doi: 10.1371/journal.pone.0270177

Figure Lengend Snippet: Description of reference strains according to ATCC.

Article Snippet: ATCC 33277D.

Techniques: Bacteria, Isolation

Journal: Journal of cell science

Article Title: Regulation of cell quiescence-proliferation balance by Ca2+-CaMKK-Akt signaling.

doi: 10.1242/jcs.253807

Figure Lengend Snippet: Fig. 2. Ca2+/CaM activity is critical in NaR cell reactivation. (A) Effect of BAPTA-AM treatment. Wild-type larvae were transferred to low [Ca2+] medium containing DMSO or 100 µM BAPTA-AM at 3 dpf. At 5 dpf, they were fixed. Because they were wild-type fish, the NaR cells were detected by in situ hybridization using a trpv6 cRNA probe. NaR cell number in each fish was manually quantified. n=21–23. (B) Tg(igfbp5a:GFP) embryos were transferred to low [Ca2+] embryo medium containing DMSO, W-7 hydrochloride (W-7, 50 µM) or calmidazolium chloride (Calmi, 1 µM) at 3 dpf. At 5 dpf, NaR cell number in each fish was quantified. n=21–23. (C,D) Changes in [Ca2+]c. Tg(igfbp5a:GCaMP7a) embryos were injected with BAC (igfbp5a:mCherry) at one-cell stage and raised until 3 dpf. They were imaged before transferring to the low [Ca2+] embryo medium (control) and at the indicated times after transfer to the low [Ca2+] stress conditions. GCaMP7a and mCherry signal intensity in the same NaR cell was measured, and the normalized change in GCaMP7a signal is expressed as percentage of the control group value. Representative images are shown in C, and quantified results are shown in D. n=6. (E,F) Changes in [Ca2+]ER. Tg(igfbp5a:GFP) embryos were injected with BAC (igfbp5a:ER-LAR-GECO-1) at one-cell stage. The injected embryos were raised until 3 dpf. They were imaged before transferring to the low [Ca2+] embryo medium (control) and at the indicated times after transfer to the low [Ca2+] stress conditions. The ER-LAR-EGCO1:GFP signal intensity ratio in the same NaR cell was measured and quantified as described in the Materials and Methods, and is expressed as a percentage of the control group value. Representative images are shown in E, and the quantified results are shown in F. n=8. Data in A,B,D and F are presented as mean±s.e.m. For images in C and E, single NaR cells are shown. *P<0.05; **P<0.01; ****P<0.0001; ns, not significant (two-tailed unpaired t-test in A; one-way ANOVA followed by Tukey’s multiple comparison test in B,D,F).

Article Snippet: Calmidazolium chloride, dantrolene and ryanodine were purchased from Tocris Bioscience (Bristol, UK).

Techniques: Activity Assay, In Situ Hybridization, Injection, Transferring, Control, Two Tailed Test, Comparison

Journal: Journal of cell science

Article Title: Regulation of cell quiescence-proliferation balance by Ca2+-CaMKK-Akt signaling.

doi: 10.1242/jcs.253807

Figure Lengend Snippet: Fig. 4. ER Ca2+ efflux regulates NaR cell reactivation via Akt. (A) Representative images of phospho-Akt immunostaining (dark color) in thapsigargin (TG)-treated and control (DMSO) wild-type fish. Images show lateral views of the zebrafish yolk sac region, with the dorsal side positioned to the top and the anterior to the left. (B) Wild-type fish were transferred to low [Ca2+] embryo media containing DMSO or 100 µM BAPTA-AM at 3 dpf. One day later, they were subjected to phospho-Akt (S473) immunostaining. The number of NaR cells positive for phospho-Akt (S473) in the yolk sac region was quantified. DMSO, n=13; BAPTA-AM, n=16. (C,D) Wild-type embryos were transferred at 3 dpf to low [Ca2+] embryo media containing (C) DMSO, W-7 hydrochloride (W-7, 50 µM) or calmidazolium chloride (Calmi, 1 µM), or (D) thapsigargin at the indicated concentrations. One day later, they were immunostained and quantified as described in B. n=11–32. One-way ANOVA followed by Tukey’s multiple comparison test. ns, no statistical difference. (E) Effect of myrAkt. One-cell-stage Tg(igfbp5a:GFP) embryos were injected with BAC(igfbp5a:myr-Akt-mCherry) or BAC(igfbp5a:mCherry) (mCherry control) and raised until 3 dpf. They were transferred to low [Ca2+] embryo medium containing DMSO or thapsigargin (0.5 µM) at 3 dpf. At 5 dpf, NaR cells labeled by both GFP and mCherry were scored following a previously established scoring system (Liu et al., 2018). NaR cells that did not divide, divided once or divided twice are scored as −,+and++, respectively. n=8–12. Data in B–D are presented as mean±s.e.m. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; ns, not significant (two-tailed unpaired t-test in B; one-way ANOVA followed by Tukey’s multiple comparison test in C,D; chi-square test in E).

Article Snippet: Calmidazolium chloride, dantrolene and ryanodine were purchased from Tocris Bioscience (Bristol, UK).

Techniques: Immunostaining, Control, Comparison, Injection, Labeling, Two Tailed Test