Journal: Cells

Article Title: Caspase-Dependent HMGB1 Release from Macrophages Participates in Peripheral Neuropathy Caused by Bortezomib, a Proteasome-Inhibiting Chemotherapeutic Agent, in Mice

doi: 10.3390/cells10102550

Figure Lengend Snippet: Involvement of HMGB1-targeted receptors in the CIPN caused by bortezomib. Bortezomib at 0.4 mg/kg or vehicle was administered i.p. on days 0, 2, 5, 7, 9, and 12. ( A , B ) Preventive ( A ) or therapeutic ( B ) effect of FPS-ZM1, TAK-242, and AMD3100, antagonists of RAGE, TLR4, and CXCR4, respectively. The mice received repeated i.p. administration of FPS-ZM1 at 1 mg/kg, TAK-242 at 3 mg/kg, AMD3100 at 8 mg/kg, or vehicle, 30 min before each dose of bortezomib ( A ), or single i.p. administration of each of them on day 14 ( B ). ( C , D ) Protein levels of RAGE, TLR4, and CXCR4 in the dorsal root ganglion ( C ) and sciatic nerves ( D ) on day 14 after the onset of bortezomib treatment. ( E , F ) Lack of preventive ( E ) and therapeutic ( F ) effects of TH1020, a TLR5 antagonist. The mice received repeated ( E ) or single ( F ) i.p. administration of TH1020 at 1 mg/kg or vehicle, in mice treated with bortezomib according to the above-mentioned schedules. Data show the mean with S.E.M for 5–6 ( A , B , F ), 5–7 ( C ), 6–7 ( D ), or 6 ( E ) mice. V, vehicle; BTZ, bortezomib; DRG, dorsal root ganglion; * p < 0.05, ** p < 0.01 vs. V ( C ) or V in V-treated mice ( A , B , E , F ). † p < 0.05, †† p < 0.01 vs. V in BTZ-treated mice.

Article Snippet: Primary antibodies were: an anti-HMGB1 rabbit polyclonal antibody (Abcam, Cambridge, UK) (1: 5000 dilution), anti-RAGE rabbit polyclonal antibody (Abcam) (1:1000 dilution), anti-TLR4 rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) (1:10000 dilution), anti-CXCR4 rabbit polyclonal antibody (Novus Biologicals, Littleton, CO, USA) (1:1000 dilution), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc.) (1:5000 dilution).

Techniques:

Journal: Cells

Article Title: Caspase-Dependent HMGB1 Release from Macrophages Participates in Peripheral Neuropathy Caused by Bortezomib, a Proteasome-Inhibiting Chemotherapeutic Agent, in Mice

doi: 10.3390/cells10102550

Figure Lengend Snippet: Scheme for bortezomib-induced caspase-dependent HMGB1 release from macrophages and CIPN development, in contrast to caspase-independent mechanisms for paclitaxel. ( A ) Inhibition of proteasome by bortezomib causes caspase-dependent apoptosis of macrophages followed by the release of HMGB1, which in turn causes neuronal sensitization via activation of RAGE and acceleration of CXCL12/CXCR4 signals, leading to CIPN. ( B ) Paclitaxel causes HMGB1 release from macrophages through activation of the ROS/p38MAPK/NF-κB pathway , independently of caspase (see D), and the extracellular HMGB1 develops CIPN in a manner dependent on RAGE and CXCR4 , as shown in the CIPN caused by bortezomib.

Article Snippet: Primary antibodies were: an anti-HMGB1 rabbit polyclonal antibody (Abcam, Cambridge, UK) (1: 5000 dilution), anti-RAGE rabbit polyclonal antibody (Abcam) (1:1000 dilution), anti-TLR4 rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) (1:10000 dilution), anti-CXCR4 rabbit polyclonal antibody (Novus Biologicals, Littleton, CO, USA) (1:1000 dilution), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc.) (1:5000 dilution).

Techniques: Inhibition, Activation Assay

Journal:

Article Title: Bicyclams, Selective Antagonists of the Human Chemokine Receptor CXCR4, Potently Inhibit Feline Immunodeficiency Virus Replication

doi:

Figure Lengend Snippet: Inhibition of binding of the anti-CXCR4 MAb to feline thymocytes in the presence (C) or absence (B) of AMD3100 at 1 μg/ml. In panel A, results obtained with an isotype control MAb are shown. The percentages of fluorescence-positive cells and the mean fluorescence intensity values are indicated in each histogram.

Article Snippet: AMD3100 had no inhibitory effect (at 100 μg/ml) on the binding of feline anti-CD4 MAb (MCA 1347; Serotec, Oxford, United Kingdom) and anti-CD8 MAb (MCA 1350; Serotec) in feline thymocytes. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 4 caption a7 Inhibition of binding of the anti-CXCR4 MAb to feline thymocytes in the presence (C) or absence (B) of AMD3100 at 1 μg/ml.

Techniques: Inhibition, Binding Assay, Fluorescence

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effects of 405 nm irradiance on chlamydial growth in HeLa cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5. ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (24 h post). Treatments are grouped based on post-hoc comparisons for convenience. The effect of 405 nm on chlamydial growth was assessed during active and persistent stages induced with penicillin ( B and C ). Growth was determined using quantitative real-time PCR to determine the ratio of chlamydial and eukaryotic housekeeping genes (16S: GAPDH respectively) 48 h post-infection on cDNA reverse transcribed from RNA. Mean ± standard deviation are plotted for the two replicated experiments. Statistical significance was determined post-hoc using a Bonferonni adjustment comparing all groups against C. trachomatis -infected HeLa cells alone (CTE); * P < 0.05, ** P < 0.005.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Real-time Polymerase Chain Reaction, Reverse Transcription, Standard Deviation

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Anti-chlamydial properties of 405 nm irradiance. ( A - C ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 without exposure to photodiodes. ( D - F ) Infected cells were exposed to 405 nm LEDs at 20 J/cm 2 promptly after infection to evaluate anti-chlamydial effects during an acute chlamydial infection. Cells were fixed and stained with dapi (blue) ( B and E ) and anti-chlamydial (green) ( C and F ) antibody 48 hours post-infection. Bar = 10μm.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Staining

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effect of 405 nm on IL-6 production in C. trachomatis -infected epithelial cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 (CTE5). ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (post-24 h). The effect of 405 nm on IL-6 production was assessed during active ( A and B ) and penicillin-induced persistent stages ( C ). Supernatants were collected and measured for IL-6 production using an ELISA. Treatments are grouped based on post-hoc comparisons for convenience. Mean ± SEM are plotted for the two replicated experiments. Statistical differences were determined post-hoc using a Bonferonni adjustment comparing all groups to C. trachomatis infected cells (CTE); *, P < 0.05; ** P < 0.005.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effect of 405 nm on CCL2 production in C. trachomatis -infected epithelial cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 (CTE5). ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (post-24 h). The effect of 405 nm on CCL2 was assessed during active ( B ) and persistent stages induced with penicillin ( C ). Supernatants were collected and measured for CCL2 production using an ELISA. Treatments are grouped based on post-hoc comparisons for convenience. Mean ± SEM are plotted for the two replicated experiments. Statistical differences were determined post-hoc using a Bonferonni adjustment comparing all groups to C. trachomatis infected cells (CTE); *, P < 0.001.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Enzyme-linked Immunosorbent Assay