Journal: BMC pharmacology & toxicology

Article Title: Narciclasine induces autophagy-mediated apoptosis in gastric cancer cells through the Akt/mTOR signaling pathway.

doi: 10.1186/s40360-021-00537-3

Figure Lengend Snippet: Fig. 4 Narciclasine promotes autophagy of gastric cancer cells. A-C, Laser confocal scanning microscopy was used to observe the formation of autolysosomes in gastric cancer cells treated with narciclasine (0.5 μM) for 24 h. D-F, Western blotting was used to detect the effect of narciclasine combined with autophagy inhibitor 3-MA (5 mM) and CQ (2.5 μM) on LC3-II and p62 protein after treatment of gastric cancer cells for 24 h. Data are shown as mean ± SD. NCS: narciclasine; 3-MA: 3-methyladenine; CQ: chloroquine

Article Snippet: The human gastric cancer cell lines BGC-823, MGC-803, GES-1, MKN28 and SGC-7901 were purchased from the Institute of Biochemistry and Cell Biology at the Chinese Academy of Sciences (Shanghai, China); Roswell Park Memorial Institute 1640 (RPMI 1640), fetal bovine serum (FBS), penicillin and streptomycin from Gibco Life Technologies (NY, US); chloroquine (CQ) and 3-methyladenine (3-MA) from Selleck Chemicals (Texas, US); insulin from Sigma-Aldrich (CA, US); and AktiVIII from Macklin Inc. (Beijing, China).

Techniques: Confocal Laser Scanning Microscopy, Western Blot

Journal: Advanced Science

Article Title: Electrochemically Synthesis of Nickel Cobalt Sulfide for High‐Performance Flexible Asymmetric Supercapacitors

doi: 10.1002/advs.201700375

Figure Lengend Snippet: The XRD patterns a) of Ni–Co–S/GF and GF. Raman spectra b) obtained for PPy/GF, Ni–Co–S/GF, GF, and PPy (inset figure (b)). XPS spectrum of Ni–Co–S/GF: c) Ni 2p, d) Co 2p, e) S 2p, (f) C 1s and inset figure (f) is XPS survey spectrum of Ni–Co–S/GF.

Article Snippet: TEM (JEOL‐2100) and high‐resolution transmission electron microscopy (HRTEM; JEOL JEM‐2010F), Raman spectra were obtained via a confocal microscope Raman spectrometer system (Witec Alpha 300; with the wavelength excitation laser of 488 nm) and XRD (Bruker Focus D8 with Cu Kα radiation, Smart lab).

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Antibacterial and Antibiofilm Activities of a Novel Synthetic Cyclic Lipopeptide against Cariogenic Streptococcus mutans UA159

doi: 10.1128/AAC.00776-17

Figure Lengend Snippet: In vitro susceptibilities of planktonic S. mutans UA159

Article Snippet: These results showed that CLP-4 is a promising agent that can effectively inhibit planktonic growth of S. mutans . table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Antimicrobial agent MIC and MBC (μg/ml) by inoculum density of: 6 × 10 5 CFU/ml 2 × 10 7 CFU/ml MIC MBC MIC MBC CLP-4 2.8 6 5 20 Erythromycin 0.016 0.6 0.062 1 Chlorhexidine dihydrochloride 1.25 3.5 1.25 5 Open in a separate window In vitro susceptibilities of planktonic S. mutans UA159 table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Streptococcus mutans strain Serotype MIC (μg/ml) MBC (μg/ml) Reference or source UA159 c 2.8 6 56 ATCC 25175 c 3.0 9 57 LM7 e 2.0 8 58 JF243 c 3.0 <5 59 568-2v-5 2.0 <5 60 764 2.0 5 C. M. Levesque 768 2.0 6 C. M. Levesque Open in a separate window S. mutans strains used in this study and their in vitro susceptibilities to CLP-4

Techniques: In Vitro

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Antibacterial and Antibiofilm Activities of a Novel Synthetic Cyclic Lipopeptide against Cariogenic Streptococcus mutans UA159

doi: 10.1128/AAC.00776-17

Figure Lengend Snippet: S. mutans strains used in this study and their in vitro susceptibilities to CLP-4

Article Snippet: These results showed that CLP-4 is a promising agent that can effectively inhibit planktonic growth of S. mutans . table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Antimicrobial agent MIC and MBC (μg/ml) by inoculum density of: 6 × 10 5 CFU/ml 2 × 10 7 CFU/ml MIC MBC MIC MBC CLP-4 2.8 6 5 20 Erythromycin 0.016 0.6 0.062 1 Chlorhexidine dihydrochloride 1.25 3.5 1.25 5 Open in a separate window In vitro susceptibilities of planktonic S. mutans UA159 table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Streptococcus mutans strain Serotype MIC (μg/ml) MBC (μg/ml) Reference or source UA159 c 2.8 6 56 ATCC 25175 c 3.0 9 57 LM7 e 2.0 8 58 JF243 c 3.0 <5 59 568-2v-5 2.0 <5 60 764 2.0 5 C. M. Levesque 768 2.0 6 C. M. Levesque Open in a separate window S. mutans strains used in this study and their in vitro susceptibilities to CLP-4

Techniques: In Vitro

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Antibacterial and Antibiofilm Activities of a Novel Synthetic Cyclic Lipopeptide against Cariogenic Streptococcus mutans UA159

doi: 10.1128/AAC.00776-17

Figure Lengend Snippet: Comparative killing kinetics of CLP-4. S. mutans UA159 cultures at a cell density of 6 × 105 CFU/ml were challenged with 5, 10, and 25 μg/ml CLP-4 under conditions of active growth in CDM supplemented with 0.5% (wt/vol) glucose (A) and against growth-arrested cells in CDM lacking any carbon source (B). Samples at time zero were enumerated prior to peptide treatment. Data shown are the means and standard deviations of three biological replicates from three independent experiments.

Article Snippet: These results showed that CLP-4 is a promising agent that can effectively inhibit planktonic growth of S. mutans . table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Antimicrobial agent MIC and MBC (μg/ml) by inoculum density of: 6 × 10 5 CFU/ml 2 × 10 7 CFU/ml MIC MBC MIC MBC CLP-4 2.8 6 5 20 Erythromycin 0.016 0.6 0.062 1 Chlorhexidine dihydrochloride 1.25 3.5 1.25 5 Open in a separate window In vitro susceptibilities of planktonic S. mutans UA159 table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Streptococcus mutans strain Serotype MIC (μg/ml) MBC (μg/ml) Reference or source UA159 c 2.8 6 56 ATCC 25175 c 3.0 9 57 LM7 e 2.0 8 58 JF243 c 3.0 <5 59 568-2v-5 2.0 <5 60 764 2.0 5 C. M. Levesque 768 2.0 6 C. M. Levesque Open in a separate window S. mutans strains used in this study and their in vitro susceptibilities to CLP-4

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Antibacterial and Antibiofilm Activities of a Novel Synthetic Cyclic Lipopeptide against Cariogenic Streptococcus mutans UA159

doi: 10.1128/AAC.00776-17

Figure Lengend Snippet: CLP-4 prevents S. mutans biofilm formation. (A) Biofilms inoculated with 2 × 107 CFU/ml were grown for 24 h in the presence of CLP-4, chlorhexidine, or erythromycin at concentrations ranging between 0.6× and 2× their respective MICs. Biofilm formation was quantified using crystal violet staining and expressed in percentage relative to untreated control. Shown are the means and standard deviations of three biological replicates from three independent experiments. *, P < 0.05; ***, P < 0.001 compared to untreated control. (B) Corresponding growth curve kinetics showing the MIC of CLP-4 on S. mutans UA159.

Article Snippet: These results showed that CLP-4 is a promising agent that can effectively inhibit planktonic growth of S. mutans . table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Antimicrobial agent MIC and MBC (μg/ml) by inoculum density of: 6 × 10 5 CFU/ml 2 × 10 7 CFU/ml MIC MBC MIC MBC CLP-4 2.8 6 5 20 Erythromycin 0.016 0.6 0.062 1 Chlorhexidine dihydrochloride 1.25 3.5 1.25 5 Open in a separate window In vitro susceptibilities of planktonic S. mutans UA159 table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Streptococcus mutans strain Serotype MIC (μg/ml) MBC (μg/ml) Reference or source UA159 c 2.8 6 56 ATCC 25175 c 3.0 9 57 LM7 e 2.0 8 58 JF243 c 3.0 <5 59 568-2v-5 2.0 <5 60 764 2.0 5 C. M. Levesque 768 2.0 6 C. M. Levesque Open in a separate window S. mutans strains used in this study and their in vitro susceptibilities to CLP-4

Techniques: Staining, Control

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Antibacterial and Antibiofilm Activities of a Novel Synthetic Cyclic Lipopeptide against Cariogenic Streptococcus mutans UA159

doi: 10.1128/AAC.00776-17

Figure Lengend Snippet: Effects of CLP-4 on preformed biofilms. S. mutans UA159 biofilms were established for 24 h and then treated with increasing concentrations (1× to 10× the MIC) of CLP-4, chlorhexidine, or erythromycin. (A) Antibiofilm activities were assessed by quantifying the cell viability of treated biofilms by colony enumeration on agar plates. The means and standard deviations of three biological replicates from three independent experiments are shown. **, P < 0.01; ***, P < 0.001 compared to untreated control. (B) Biofilms treated with 10× the MICs for each antimicrobial were fluorescently labeled using the LIVE/DEAD BacLight viability stain and visualized by confocal laser scanning microscopy. Shown are the top-down three-dimensional (3D) volume rendering of biofilms at a total magnification of ×400. Bottom images represent optical planes in the xz, and vertical thin images represent yz dimensions. Membrane-compromised bacteria are stained red with propidium iodide, while intact bacteria are stained green with SYTO 9. Areas highlighted by dashed lines indicate regions of interest (ROIs) viewed at a higher magnification. Dimensions shown are 387.5 μm by 387.5 μm by 16 μm. (C) ROIs are presented at ×2,300 magnification. Dimensions shown are 68.1 μm by 68.1 μm by 16 μm.

Article Snippet: These results showed that CLP-4 is a promising agent that can effectively inhibit planktonic growth of S. mutans . table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Antimicrobial agent MIC and MBC (μg/ml) by inoculum density of: 6 × 10 5 CFU/ml 2 × 10 7 CFU/ml MIC MBC MIC MBC CLP-4 2.8 6 5 20 Erythromycin 0.016 0.6 0.062 1 Chlorhexidine dihydrochloride 1.25 3.5 1.25 5 Open in a separate window In vitro susceptibilities of planktonic S. mutans UA159 table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Streptococcus mutans strain Serotype MIC (μg/ml) MBC (μg/ml) Reference or source UA159 c 2.8 6 56 ATCC 25175 c 3.0 9 57 LM7 e 2.0 8 58 JF243 c 3.0 <5 59 568-2v-5 2.0 <5 60 764 2.0 5 C. M. Levesque 768 2.0 6 C. M. Levesque Open in a separate window S. mutans strains used in this study and their in vitro susceptibilities to CLP-4

Techniques: Control, Labeling, Staining, Confocal Laser Scanning Microscopy, Membrane, Bacteria

Journal: The FASEB Journal

Article Title: Leptin regulates parathyroid hormone secretion through CaSR ‐ ERK1 /2 signaling

doi: 10.1096/fj.202403141R

Figure Lengend Snippet: Reduced serum PTH levels in db/db mice, with no change in PTH and CasR mRNA levels. (A, B) Serum PTH levels in db/− and db/db mice at 4 (A) and 7 (B) months of age. (C, D) Serum calcium and blood urea nitrogen (BUN) levels at 7 months of age. (E, F) PTH and CaSR mRNA levels analyzed by qRT‐PCR of RNA extracted from thyroparathyroid glands of db/− and db/db mice at 4 (E) and 7 (F) months of age. * p < .05; *** p < .001.

Article Snippet: Immunostaining was conducted overnight at 4°C using the following primary antibodies diluted in Cas block (Zymed Laboratories, San Francisco, CA): PTH at a dilution of 1:300 (Bio‐Rad, Hercules, CA, RRID:AB 7170-6216); CaSR at a dilution of 1:1500 (Novus Biologicals, Littleton, CO, RRID:AB 7170-6216), klotho at a dilution of 1:100 (Labome, Lambertville, NJ, RRID:AB KLO001); FGFR1 at a dilution of 1:200 (Santa Cruz Biotechnology Dallas, TX, RRID:AB Flg (C-15): sc-121).

Techniques: Quantitative RT-PCR

Journal: The FASEB Journal

Article Title: Leptin regulates parathyroid hormone secretion through CaSR ‐ ERK1 /2 signaling

doi: 10.1096/fj.202403141R

Figure Lengend Snippet: Reduced PTH content in parathyroid sections from db/db mice at 4 months of age. Parathyroid glands form db/− and db/db mice were extracted at 4 months of age, paraffin embedded and analyzed by IF microscopy. (A) Representative staining for PTH (blue), nuclear sytox (green) and the merged image. (B) Quantification of staining intensity for PTH. (C) Representative staining for CaSR (blue), klotoho (red) and nuclear sytox (green) and their quantification (D, E). (F) Reprisentative staining for FGFR1 (red), nuclear sytox (green) and the merged image and quantification (G). Quantification was performed using Image J quantification program. * p < .05.

Article Snippet: Immunostaining was conducted overnight at 4°C using the following primary antibodies diluted in Cas block (Zymed Laboratories, San Francisco, CA): PTH at a dilution of 1:300 (Bio‐Rad, Hercules, CA, RRID:AB 7170-6216); CaSR at a dilution of 1:1500 (Novus Biologicals, Littleton, CO, RRID:AB 7170-6216), klotho at a dilution of 1:100 (Labome, Lambertville, NJ, RRID:AB KLO001); FGFR1 at a dilution of 1:200 (Santa Cruz Biotechnology Dallas, TX, RRID:AB Flg (C-15): sc-121).

Techniques: Microscopy, Staining

Journal: The FASEB Journal

Article Title: Leptin regulates parathyroid hormone secretion through CaSR ‐ ERK1 /2 signaling

doi: 10.1096/fj.202403141R

Figure Lengend Snippet: Schematic representation of interactions between adipose tissue, parathyroid glands, and kidneys mediated by leptin, PTH, and 1,25(OH) 2 D 3 . Leptin, predominantly secreted by adipocytes, directly stimulates PTH secretion by inhibiting the CasR/pERK1/2 signaling pathway, thereby lifting the CasR‐mediated suppression of PTH secretion by the parathyroid glands. Elevated PTH levels, in turn, may affect leptin production by adipocytes. ⁷ , ¹⁷ Additionally, the parathyroid glands themselves produce leptin, further integrating these pathways. PTH stimulates renal synthesis of 1,25(OH) 2 D 3 (1,25D) which suppresses PTH secretion and contributes to calcium and phosphate homeostasis. Leptin reduces 1,25D, thus promoting PTH secretion indirectly by decreasing the inhibitory feedback exerted by 1,25D on the parathyroid glands. This intricate regulatory network links leptin to mineral homeostasis through PTH, CaSR, and 1,25D.

Article Snippet: Immunostaining was conducted overnight at 4°C using the following primary antibodies diluted in Cas block (Zymed Laboratories, San Francisco, CA): PTH at a dilution of 1:300 (Bio‐Rad, Hercules, CA, RRID:AB 7170-6216); CaSR at a dilution of 1:1500 (Novus Biologicals, Littleton, CO, RRID:AB 7170-6216), klotho at a dilution of 1:100 (Labome, Lambertville, NJ, RRID:AB KLO001); FGFR1 at a dilution of 1:200 (Santa Cruz Biotechnology Dallas, TX, RRID:AB Flg (C-15): sc-121).

Techniques:

Journal: The FASEB Journal

Article Title: Leptin regulates parathyroid hormone secretion through CaSR ‐ ERK1 /2 signaling

doi: 10.1096/fj.202403141R

Figure Lengend Snippet: Leptin increases PTH secretion in microdissected parathyroid glands in culture while decreasing CasR and c‐fos gene expression. (A) Parathyroid glands from control mice with intact leptin regeptor and parathyroid‐specific td‐tomato ‐expression were identified (dotted lines) and excised using fluorescent‐guided microdissection. Glands were cultured in growth medium supplemented with leptin (1 μg/mL) or vehicle. (B) PTH accumulated in the growth medium at 3 h. (C) qRT‐PCR analysis of PTH and CaSR mRNA levels performed on RNA extracted from the glands in A, after 24 h incubation. (D) qRT‐PCR analysis of c‐fos mRNA levels performed on RNA extracted after 3 h incubation. (E) Parathyroid glands were incubated with the calcimimatic R568 (1 μM) in the presence or absence of leptin, and PTH accumulated in the growth medium measured at 3 h. * p < .05, ** p < .01.

Article Snippet: Immunostaining was conducted overnight at 4°C using the following primary antibodies diluted in Cas block (Zymed Laboratories, San Francisco, CA): PTH at a dilution of 1:300 (Bio‐Rad, Hercules, CA, RRID:AB 7170-6216); CaSR at a dilution of 1:1500 (Novus Biologicals, Littleton, CO, RRID:AB 7170-6216), klotho at a dilution of 1:100 (Labome, Lambertville, NJ, RRID:AB KLO001); FGFR1 at a dilution of 1:200 (Santa Cruz Biotechnology Dallas, TX, RRID:AB Flg (C-15): sc-121).

Techniques: Gene Expression, Control, Expressing, Laser Capture Microdissection, Cell Culture, Quantitative RT-PCR, Incubation

Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Detecting Purinosome Metabolon Formation with Fluorescence Microscopy

doi: 10.1007/978-1-4939-7759-8_17

Figure Lengend Snippet: Colocalization of endogenous FGAMS and GART for visualization of purinosomes by immunofluorescence. Purine-depleted HeLa cells were fixed and permeabilized prior to being probed for with FGAMS rabbit polyclonal antibody and GART mouse monoclonal antibody. Fluorescently labeled secondary antibodies CF488A-conjugated donkey anti-rabbit and CF568-conjugated donkey anti-mouse were used to visualize the expression and localization of FGAMS and GART, respectively. A representative image of an individual cell was captured using a 100× oil objective on an Olympus Fluoview 1000 confocal laser scanning microscope. Sequential imaging of CF488A and CF568 showed colocalization of (a) FGAMS with (b) GART as represented by the yellow puncta present in (c) the merged image. Scale bar: 10 μm

Article Snippet: Primary antibody solution: 1:500 dilution of PFAS rabbit polyclonal antibody (Bethyl Laboratories) and 1:1000 dilution of GART mouse monoclonal antibody (Novus Biologicals) prepared in blocking buffer.

Techniques: Immunofluorescence, Labeling, Expressing, Laser-Scanning Microscopy, Imaging