Journal: PLoS ONE

Article Title: Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes

doi: 10.1371/journal.pone.0192680

Figure Lengend Snippet: 1 day cultured monocytes were collected for total RNA extraction and iOPN expression, and culture supernatants for OPN ELISA; quantitative real-time PCR was performed for OPN gene expression. iOPN expression was measured by flow cytometry. MGBG decreased OPN gene expression and sOPN level in a dose-dependent manner. MGBG slightly decreased iOPN expression. The data were normalized against that of untreated controls. The average sOPN level for 1 day untreated monocytes was approximately 3 ng/ml. The iOPN geometric mean florescence of 1 day untreated cells varies among individuals ranging from 209–689. The OPN RNA level was calculated from β-actin normalized Ct difference of 1D cultured cells versus cells at isolation. The OPN Ct difference of 1 day untreated monocytes also varies among individuals ranging from 1.7 to 9.9. ED50 of MGBG on OPN cytokine level and gene expression on day 1: ~ 0.1 μM, n = 4, means and SEM.

Article Snippet: MGBG (Ash Stevens, Riverview, MI) and spermine (SPM) (Sigma-Aldrich, St. Louis, MO) were added where specified.

Techniques: Cell Culture, RNA Extraction, Expressing, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Gene Expression, Flow Cytometry, Isolation

Journal: PLoS ONE

Article Title: Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes

doi: 10.1371/journal.pone.0192680

Figure Lengend Snippet: (A). MGBG significantly inhibited sOPN production in monocytes. The degree of MGBG inhibition on sOPN was related to time of exposure. Isolated human monocytes were cultured at 0.1 million cells per ml with various concentrations of MGBG for 16 hr, 1, 2 and 3 days. MGBG showed potent inhibition on monocyte sOPN production with IC50 < 0.1 μM on day 3. Data were presented as percentage of untreated controls. The average sOPN levels of untreated monocytes were approximately 110, 130, 670 and 5300 pg/ml at 16 hr, 1, 2, and 3 days, respectively. The inhibition is independent of cytotoxicity (e.g. little cell death at 16 hours and day 1). The data is normalized to number of live cells in the culture. n = 2. (B). MGBG showed limited inhibitory effect on sOPN production in mature macrophages. Isolated monocytes were cultured to allowed to differentiate into macrophages for 6 days prior to drug treatment. Fresh media was replaced and cells were treated with 10 μM MGBG for 3 days. n = 4, means and SEM.

Article Snippet: MGBG (Ash Stevens, Riverview, MI) and spermine (SPM) (Sigma-Aldrich, St. Louis, MO) were added where specified.

Techniques: Inhibition, Isolation, Cell Culture

Journal: PLoS ONE

Article Title: Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes

doi: 10.1371/journal.pone.0192680

Figure Lengend Snippet: Isolated human monocytes were cultured in suspension with or without MGBG treatment. CD16 expression was measured by flow cytometry. (A). Monocytes differentiate spontaneously in culture. CD16 expression level increases in monocyte culture. n = 4, means and SEM. (B). MGBG inhibited CD16 expression in 1 day cultured monocytes in a dose-dependent manner. The average CD16 geometric mean fluorescence of 1 day untreated monocytes was measured at 108 units. n = 4, means and SEM. (C). MGBG inhibited BrdU incorporation in cultured monocytes. Isolated human monocytes were cultured for 3–6 days for BrdU incorporation using a FITC BrdU flow kit. The average percentage of BrdU incorporated untreated monocytes was measured at 4.7%. MGBG inhibited monocyte BrdU uptake in a dose-dependent manner. n = 5, means and SEM.

Article Snippet: MGBG (Ash Stevens, Riverview, MI) and spermine (SPM) (Sigma-Aldrich, St. Louis, MO) were added where specified.

Techniques: Isolation, Cell Culture, Suspension, Expressing, Flow Cytometry, Fluorescence, BrdU Incorporation Assay

Journal: PLoS ONE

Article Title: Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes

doi: 10.1371/journal.pone.0192680

Figure Lengend Snippet: Monocytes were isolated from PBMCs using CD14 microbeads. Cells were cultured for 1, 3, and 6 days with various concentrations of MGBG, and collected for CD16 expression measurement using flow cytometry. MGBG showed greater inhibitory effect on CD 16 expression in 1 and 3 day monocytes than that in differentiated macrophages. The average CD16 geometric mean fluorescence was measured at 108, 448, and 249 for 1, 3, and 6 day untreated cells, respectively. n = 4, means and SEM.

Article Snippet: MGBG (Ash Stevens, Riverview, MI) and spermine (SPM) (Sigma-Aldrich, St. Louis, MO) were added where specified.

Techniques: Isolation, Cell Culture, Expressing, Flow Cytometry, Fluorescence

Journal: PLoS ONE

Article Title: Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes

doi: 10.1371/journal.pone.0192680

Figure Lengend Snippet: PBMCs were cultured for 3 days with or without treatment. Cells were cultured with 10 μM MGBG and various concentrations of SPM. (A). MGBG significantly decreased sOPN; SPM reversed the MGBG inhibitory effects on sOPN. The average sOPN level of 3 day untreated and 10 μM MGBG treated cells was 30 and 2 ng/ml, respectively. n = 6, means and SEM. (B). MGBG inhibited monocyte CD16 expression; SPM reversed the MGBG inhibitory effects on CD16 expression. Cultured PBMCs were double stained with CD14 and CD16 antibodies for flow cytometry analysis. The average CD16 geometric mean in untreated and MGBG treated CD14+ monocytes were measured at 510 and 249, respectively. n = 3, means and SEM. Data was presented as a percentage of inhibition of MGMG treatment only.

Article Snippet: MGBG (Ash Stevens, Riverview, MI) and spermine (SPM) (Sigma-Aldrich, St. Louis, MO) were added where specified.

Techniques: Cell Culture, Expressing, Staining, Flow Cytometry, Inhibition

Journal: PLoS ONE

Article Title: Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes

doi: 10.1371/journal.pone.0192680

Figure Lengend Snippet: Isolated human monocytes were cultured to differentiate and induced to M1 or M2 macrophages using 50 ng/ml INF-γ and 1 μg/ml LPS or 50 ng/ml IL4 for 5–6 days with or without 0.4 μM MGBG. Cells were cultured and evaluated by flow cytometry at day 5 or 6. Mono: Baseline expression of cell surface markers in 4 hr cultured monocytes (% of all monocytes); Mac: Cultured in normal media -/+ 0.4 μM MGBG; M1: Cultured in M1 polarizing media -/+ 0.4 μM MGBG; M2: Cultured in M2 polarizing media -/+ 0.4 μM MGBG. MGBG inhibited monocyte differentiation in both M1 and M2 macrophages. n = 2.

Article Snippet: MGBG (Ash Stevens, Riverview, MI) and spermine (SPM) (Sigma-Aldrich, St. Louis, MO) were added where specified.

Techniques: Isolation, Cell Culture, Flow Cytometry, Expressing

Journal: Journal of neurovirology

Article Title: An oral form of methylglyoxal-bis-guanylhydrazone reduces monocyte activation and traffic to the dorsal root ganglia in a primate model of HIV-peripheral neuropathy

doi: 10.1007/s13365-017-0529-9

Figure Lengend Snippet: SIV+ rhesus macaques used in this study and DRG pathology.

Article Snippet: Nine animals received 30mg/kg of MGBG (provided by Pathologica, LLC; formulated as flavored syrup by Wedgewood Pharmacy, Swedesboro, NJ) daily beginning at 21 dpi.

Techniques: Control

Journal: Journal of neurovirology

Article Title: An oral form of methylglyoxal-bis-guanylhydrazone reduces monocyte activation and traffic to the dorsal root ganglia in a primate model of HIV-peripheral neuropathy

doi: 10.1007/s13365-017-0529-9

Figure Lengend Snippet: The numbers of CD68+ (A-C), CD163+ (D-F), MAC387+ (G-I), BrdU+ (J-L), and CD3+ (M-O) cells per mm2 of tissue were counted in DRG from control animals and MGBG-treated animals. Representative images of DRG from control animals (A, D, G, J, M) and MGBG-treated animals (B, E, H, K, N) are shown. Data are shown as mean ± SEM (C, F, I, L, O). Groups were compared with a Mann-Whitney T-test. *P<0.05; **P<0.01; ****P<0.0001.

Article Snippet: Nine animals received 30mg/kg of MGBG (provided by Pathologica, LLC; formulated as flavored syrup by Wedgewood Pharmacy, Swedesboro, NJ) daily beginning at 21 dpi.

Techniques: Control, MANN-WHITNEY

Journal: Journal of neurovirology

Article Title: An oral form of methylglyoxal-bis-guanylhydrazone reduces monocyte activation and traffic to the dorsal root ganglia in a primate model of HIV-peripheral neuropathy

doi: 10.1007/s13365-017-0529-9

Figure Lengend Snippet: IENFD was measured at 20 dpi and at necropsy in control (A) and in MGBG-treated (B) groups. Changes in IENFD between time points assessed with a Wilcoxon matched-pairs signed rank test. A p value less than 0.05 was considered significant.

Article Snippet: Nine animals received 30mg/kg of MGBG (provided by Pathologica, LLC; formulated as flavored syrup by Wedgewood Pharmacy, Swedesboro, NJ) daily beginning at 21 dpi.

Techniques: Control

Journal: Journal of acquired immune deficiency syndromes (1999)

Article Title: Direct Targeting of Macrophages with Methylglyoxal-Bis-Guanylhydrazone Decreases SIV-Associated Cardiovascular Inflammation and Pathology

doi: 10.1097/QAI.0000000000001297

Figure Lengend Snippet: Animals Used in the Study.

Article Snippet: Of these, 11 received MGBG (30 mg/kg) (provided by Pathologica, LLC; formulated as syrup by Wedgewood Pharmacy, Swedesboro, NJ) orally, once daily beginning at 21 days post infection (dpi), when significant CNS and cardiovascular inflammation is known to occur in this model. 24 , 42 Eight received on oral placebo-syrup control at the same time point.

Techniques:

Journal: Journal of acquired immune deficiency syndromes (1999)

Article Title: Direct Targeting of Macrophages with Methylglyoxal-Bis-Guanylhydrazone Decreases SIV-Associated Cardiovascular Inflammation and Pathology

doi: 10.1097/QAI.0000000000001297

Figure Lengend Snippet: (A) Sections of carotid artery from MGBG treated (n=9) and placebo control (n=6) SIV-infected CD8+ T-lymphocyte rhesus macaques were immunohistochemically stained with antibodies recognizing CD163+, CD68+, CD206+, and MAC387+ macrophages. MGBG treatment resulted in a trend of decreased numbers of CD163+ (2.19-fold), CD68+ (1.86-fold), MAC387+ (2.12-fold), and CD206+ (2.31-fold) macrophages present in the carotid artery when compared to placebo controls. Statistical analysis was performed using a non-parametric Mann-Whitney t-test with significance accepted at p<0.05. (B) Sections of cardiac tissue from SIV- (n=6) MGBG treated (n=11) and placebo controls (n=8) rhesus macaques were immunohistochemically stained with the same antibodies. MGBG treated animals had significantly decreased numbers of CD163+ (2.07-fold), CD68+ (1.61-fold), and MAC387+ (1.95-fold) and CD206+ (1.62-fold) macrophages when compared to placebo controls. There was no difference in numbers of CD163+, MAC387+, and CD206+ macrophages when comparing uninfected and MGBG treated animals. For cardiac tissues statistical analysis was performed between uninfected animals, SIV-infected placebo controls, and SIV-infected MGBG treated animals using analysis of variance (ANOVA). If the ANOVA was significant (p<0.05) between two groups, a posthoc non-parametric Mann-Whitney t-test was performed. * p<0.05, ** p<0.01.

Article Snippet: Of these, 11 received MGBG (30 mg/kg) (provided by Pathologica, LLC; formulated as syrup by Wedgewood Pharmacy, Swedesboro, NJ) orally, once daily beginning at 21 days post infection (dpi), when significant CNS and cardiovascular inflammation is known to occur in this model. 24 , 42 Eight received on oral placebo-syrup control at the same time point.

Techniques: Control, Infection, Staining, MANN-WHITNEY

Journal: Journal of acquired immune deficiency syndromes (1999)

Article Title: Direct Targeting of Macrophages with Methylglyoxal-Bis-Guanylhydrazone Decreases SIV-Associated Cardiovascular Inflammation and Pathology

doi: 10.1097/QAI.0000000000001297

Figure Lengend Snippet: (A) Sections of carotid artery were stained with a Verhoeff Van Gieson elastic stain in order to measure the carotid artery intima-media thickness (cIMT). The cIMT was measured using calipers and ImageJ Analysis Software from 10 random non-overlapping fields of view with average cIMT calculated and expressed as plus or minus the standard error of the mean (SEM). (B) There was a significant 1.49-fold decrease in cIMT when comparing placebo control animals to MGBG treated animals. Statistical analysis was performed using a non-parametric Mann-Whitney t-test comparing placebo controls to MGBG treated animals with significance accepted at p<0.05. (C) Using a Spearman rank correlation analysis, in the carotid artery, there was a positive correlation between increased numbers of CD68+ (r=0.52) and MAC387+ (r=0.42) macrophages and increasing cIMT. There was no correlation between numbers of CD163+ and CD206+ macrophages and cIMT. * p<0.05. Scale bar equals 50 μm.

Article Snippet: Of these, 11 received MGBG (30 mg/kg) (provided by Pathologica, LLC; formulated as syrup by Wedgewood Pharmacy, Swedesboro, NJ) orally, once daily beginning at 21 days post infection (dpi), when significant CNS and cardiovascular inflammation is known to occur in this model. 24 , 42 Eight received on oral placebo-syrup control at the same time point.

Techniques: Staining, Software, Control, MANN-WHITNEY

Journal: Journal of acquired immune deficiency syndromes (1999)

Article Title: Direct Targeting of Macrophages with Methylglyoxal-Bis-Guanylhydrazone Decreases SIV-Associated Cardiovascular Inflammation and Pathology

doi: 10.1097/QAI.0000000000001297

Figure Lengend Snippet: (A) Collagen, a marker of fibrosis, in cardiac tissues was measured by staining cardiac tissue with a trichrome stain. ImageJ Analysis software was used to determine the percent collagen (blue dye) per total tissue area from 20 200× non-overlapping fields of view with the average percent collagen per total tissue area calculated for each animal. (B) There was a significant 2.05-fold decrease in the percent collage per total tissue area when comparing placebo controls to MGBG treated animals. There was no significant difference between the percent collagen per total tissue area between uninfected and MGBG treated animals. Statistical analysis was performed using analysis of variance (ANOVA). If the ANOVA was significant (p<0.05) between two groups, a posthoc non-parametric Mann-Whitney t-test was performed. (C) Using Spearman rank correlation analysis, we found positive correlations between increasing numbers of CD163+ (r=0.69), CD68+ (r=0.63), MAC387+ (r=0.53) and CD206+ (r=0.54) macrophages and increasing percent collagen per total tissue area. * p<0.05, ** p<0.01. Scale bar equals 20 μm.

Article Snippet: Of these, 11 received MGBG (30 mg/kg) (provided by Pathologica, LLC; formulated as syrup by Wedgewood Pharmacy, Swedesboro, NJ) orally, once daily beginning at 21 days post infection (dpi), when significant CNS and cardiovascular inflammation is known to occur in this model. 24 , 42 Eight received on oral placebo-syrup control at the same time point.

Techniques: Marker, Staining, Software, MANN-WHITNEY

Journal: Plants

Article Title: Modulation of Organogenesis and Somatic Embryogenesis by Ethylene: An Overview

doi: 10.3390/plants10061208

Figure Lengend Snippet: Effect of ethylene modulators, mutants, or transgenic lines on different regeneration systems of diverse plant species. Ethylene modulators/mutants or transgenic lines in the effect column showed an increase (↑) or a decrease (↓) in the respective parameter when compared to control.

Article Snippet: Interestingly, in Medicago truncatula , a species from the same genus as alfalfa ( Medicago sativa ), treatments with the ethylene precursors ACC and MGBG increased the number of somatic embryos developed per embryogenic callus, while silver nitrate and AVG greatly decreased the somatic embryo formation [ ].

Techniques: Transgenic Assay, Mutagenesis, Concentration Assay, Produced, Expressing, Serial Time-encoded Amplified Microscopy