Journal: European Thyroid Journal

Article Title: Alterations in gene expression associated with invasion of RAS-mutant thyroid tumors and their potential diagnostic and therapeutic utility

doi: 10.1530/ETJ-25-0022

Figure Lengend Snippet: Effect of chemical inhibition of CA12 on proliferation, migration, and invasion of RAS -mutant cells. (A) Proliferation of EAM306 and CUTC61 cells after treatment with U-104 at concentrations of 10, 50, and 100 μM for 24, 48, and 72 h with relative proliferation compared to cells treated with DMSO. (B) Proliferation of EAM306 and CUTC61 cells after treatment with acetazolamide (ACZ) at concentrations of 1, 10, 100, or 1,000 μM for 24, 48, and 72 h with relative proliferation compared to cells treated with DMSO. (C and D) Wound healing in scratch assay of EAM306 and CUTC61 cells after treatment with U-104 at concentrations of 10, 50, and 100 μM or ACZ at concentrations of 1, 10, 100, or 1,000 μM at 24, 48, and 72 h after scratch. Relative wound closure is compared to cells treated with DMSO. (E and F) Invasion of EAM306 and CUTC61 cells across a Matrigel insert after treatment with U-104 at concentrations of 10, 50, and 100 μM or ACZ at concentrations of 1, 10, 100, or 1,000 μM for 24 h before seeding on the Matrigel insert. Relative invasion is compared to cells treated with DMSO.

Article Snippet: CA12 inhibitors, Acetazolamide (ACZ) (cat# S4506) and U-104 (cat #S2866) were purchased from Selleck Chemicals (USA).

Techniques: Inhibition, Migration, Mutagenesis, Wound Healing Assay

Journal: bioRxiv

Article Title: Hypoxia induced carbonic anhydrase mediated dorsal horn sensory neuron activation and induction of neuropathic pain

doi: 10.1101/2021.06.08.447539

Figure Lengend Snippet: Inhibition of Carbonic Anhydrase 7 attenuates hypoxia induced pain hypersensitivity [A] In vitro lumbar spinal cord slices were treated with either Vehicle or 1mM DMOG for 24hrs to determine protein expression of HIF1 α and CA7 (Representative western blots), with [B] DMOG treatment increasing expression of [B] HIF1 α and [C] CA7 compared with vehicle treatment (**P<0.01, Unpaired T Test, n=5 per group). In C57Bl6 mice, lumbar spinal cord protein lysate samples ([D] Representative western blots) 24hrs following intrathecal delivery of 1mM DMOG demonstrated an increase in protein expression of [E] HIF1 α and [F] CA7 compared with vehicle treated animals (*P<0.05, ***P<0.001, Unpaired T Test, n=3 per group). [G] Intraperitoneal treatment of acetazolamide (ACZ) led to attenuation of DMOG induced reduction in mechanical withdrawal thresholds and [H] heat withdrawal latencies (**P<0.01, ***P<0.001 Two Way ANOVA with Tukey’s multiple comparison, n=5 per group). ACZ treatment led to nociceptive withdrawal thresholds returning to baseline values and comparable to vehicle treated animals. [I] Intraperitoneal ACZ inhibited heat hypersensitivity in VEGFR2 scECKO mice with heat withdrawal latencies increased compared with VEGFR2 scECKO mice treated with vehicle (***P<0.001 Two Way ANOVA with Tukey’s multiple comparison, n=5 per group).

Article Snippet: Acetazolamide was delivered via intraperitoneal injection (40 mg/kg, in 1%DMSO in PBS, Tocris).

Techniques: Inhibition, In Vitro, Expressing, Western Blot, Comparison

Journal: Antioxidants

Article Title: Thioamide Compound H0802 Enhances Hypoxia Tolerance by Mimicking Hypoxia-Adaptive Reprogramming of Glucose and Oxygen Metabolism

doi: 10.3390/antiox15050525

Figure Lengend Snippet: H0802 improved hypoxia tolerance in mice. ( A ) Screening workflow. ( B ) Screening results of 44 compounds. ( C ) Survival curves of mice in a sealed hypoxia chamber ( n = 10). ( D ) Residual oxygen concentration in sealed chambers at the time of death ( n = 5). ( E ) Survival rates in the acute hypobaric hypoxia chamber experiment. ( n = 8). Data are presented as the mean ± SEM. *** p < 0.001, **** p < 0.0001 vs. control; ns, not significant. ( F , G ) Effects of H0802 on body weight ( F ) and food intake ( G ) in mice (100 mg/kg/day, i. g. once a day, 14-day, n = 5).

Article Snippet: H0802 (75 mg/kg) and acetazolamide (100 mg/kg, MCE, Cat. #HY-B0782) were administered by oral gavage daily for 3 consecutive days.

Techniques: Concentration Assay, Control

Journal: Antioxidants

Article Title: Thioamide Compound H0802 Enhances Hypoxia Tolerance by Mimicking Hypoxia-Adaptive Reprogramming of Glucose and Oxygen Metabolism

doi: 10.3390/antiox15050525

Figure Lengend Snippet: H0802- and hypoxia-treatment mice share common key changes in gene expression. ( A ) Workflow of RNA-seq analysis. ( B ) The heatmap of the shared gene between H0802- and hypoxia- treatment. ( C ) Volcano plots of differentially expressed genes (DEGs). ( D ) Correlation and functional enrichment analyses of DEGs. ( E ) GSEA for the 3 h H0802-treated group. ( F ) GO analysis of upregulated genes in the H0802-treated group. ( G ) GO analysis of downregulated genes in the H0802-treated group. ( H ) Validation results of the overlap between H0802-induced and hypoxia-induced DEGs. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. control; ns, not significant.

Article Snippet: H0802 (75 mg/kg) and acetazolamide (100 mg/kg, MCE, Cat. #HY-B0782) were administered by oral gavage daily for 3 consecutive days.

Techniques: Gene Expression, RNA Sequencing, Functional Assay, Biomarker Discovery, Control

Journal: Antioxidants

Article Title: Thioamide Compound H0802 Enhances Hypoxia Tolerance by Mimicking Hypoxia-Adaptive Reprogramming of Glucose and Oxygen Metabolism

doi: 10.3390/antiox15050525

Figure Lengend Snippet: H0802 promotes the stabilization and transcriptional activity of HIF-1α and HIF-2α. ( A ) H0802 enhances HIF-1α and HIF-2α proteins’ stability. ( B ) Quantification of HIF-1α and HIF-2α protein levels. ( C , D ) CHX chase assays in HepG2 cells. ( E ) The impacts of H0802 (40 μM) and roxadustat (10 μM) on endogenous HIF-1α protein ubiquitination. ( F ) QPCR analysis of HIF genes’ expression in HepG2 cells. Statistical significance: two-way ANOVA followed by Tukey’s post hoc test ( B , D , F ); * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. control. ns, not significant.

Article Snippet: H0802 (75 mg/kg) and acetazolamide (100 mg/kg, MCE, Cat. #HY-B0782) were administered by oral gavage daily for 3 consecutive days.

Techniques: Activity Assay, Ubiquitin Proteomics, Expressing, Control

Journal: Antioxidants

Article Title: Thioamide Compound H0802 Enhances Hypoxia Tolerance by Mimicking Hypoxia-Adaptive Reprogramming of Glucose and Oxygen Metabolism

doi: 10.3390/antiox15050525

Figure Lengend Snippet: H0802 regulates glucose metabolism and promotes glucose uptake. ( A ) Effects of hypoxia (10% O 2 ) or H0802 (100 mg/kg) on blood glucose levels in mice, n = 9. ( B ) Time-course changes in blood glucose levels in mice over 24 h following H0802 (100 mg/kg) treatment, n = 3. ( C ) Hepatic glycogen staining in mice treated with H0802 (100 mg/kg), scale bar: 100 μm, n = 3. ( D ) Effects of H0802 treatment on hepatic glycogen content in mice under normoxic or hypoxic conditions, n = 5. ( E ) Effects of H0802 treatment on mRNA expression of Slc2a1 in the brain tissue of mice under normoxic or hypoxic conditions, n = 6. ( F ) Glucose uptake in PC12 cells treated with DMSO or H0802 under normoxia or hypoxia. Data are shown as the mean ± SEM. p < 0.001. ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. control; ns, not significant.

Article Snippet: H0802 (75 mg/kg) and acetazolamide (100 mg/kg, MCE, Cat. #HY-B0782) were administered by oral gavage daily for 3 consecutive days.

Techniques: Staining, Expressing, Control

Journal: Antioxidants

Article Title: Thioamide Compound H0802 Enhances Hypoxia Tolerance by Mimicking Hypoxia-Adaptive Reprogramming of Glucose and Oxygen Metabolism

doi: 10.3390/antiox15050525

Figure Lengend Snippet: H0802 regulated oxygen consumption in mice. ( A ) Oxygen consumption in vehicle- or H0802-treated (75 mg/kg) mice. Values represent the mean ± SEM. ( B ) Carbon dioxide (CO 2 ) production in vehicle- or H0802-treated mice. ( C ) Respiratory quotient (RQ) in vehicle- or H0802-treated mice. ( D ) Energy expenditure (EE) in vehicle- or H0802-treated mice. Statistical significance was assessed using an unpaired two-sided Student’s t -test ( A – D ), n = 8. * p < 0.05 and ** p < 0.01 vs. control.

Article Snippet: H0802 (75 mg/kg) and acetazolamide (100 mg/kg, MCE, Cat. #HY-B0782) were administered by oral gavage daily for 3 consecutive days.

Techniques: Control

Journal: Antioxidants

Article Title: Thioamide Compound H0802 Enhances Hypoxia Tolerance by Mimicking Hypoxia-Adaptive Reprogramming of Glucose and Oxygen Metabolism

doi: 10.3390/antiox15050525

Figure Lengend Snippet: H0802 alleviates hypoxia-induced lung inflammation. ( A ) Representative hematoxylin–eosin (H&E) staining images. Scale bars: 5 mm (low-magnification overview) and 300 μm (high-magnification detail). ( B ) Quantification of lung injury scores. ( C ) Inhibitory effect of H0802 and acetazolamide on serum inflammatory cytokines (TNF-α, IL-6, IL-1β). ( D ) Differential gene expression analysis under hypoxia and H0802 treatment. ( E ) Validation results for Panel ( D ). Data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. control. ns, not significant.

Article Snippet: H0802 (75 mg/kg) and acetazolamide (100 mg/kg, MCE, Cat. #HY-B0782) were administered by oral gavage daily for 3 consecutive days.

Techniques: Staining, Gene Expression, Biomarker Discovery, Control

Journal: Pharmaceutics

Article Title: Comparative Analysis of Morphological and Release Profiles in Ocular Implants of Acetazolamide Prepared by Electrospinning

doi: 10.3390/pharmaceutics13020260

Figure Lengend Snippet: FTIR spectra of MD implants compared with PCL, Lutrol F127 and acetazolamide.

Article Snippet: Acetazolamide, purity 99%, was purchased from Alfa Aesar (Heysham, UK).

Techniques:

Journal: Pharmaceutics

Article Title: Comparative Analysis of Morphological and Release Profiles in Ocular Implants of Acetazolamide Prepared by Electrospinning

doi: 10.3390/pharmaceutics13020260

Figure Lengend Snippet: FTIR spectra of EDW implants compared with PCL, Lutrol F127 and acetazolamide.

Article Snippet: Acetazolamide, purity 99%, was purchased from Alfa Aesar (Heysham, UK).

Techniques:

Journal: Pharmaceutics

Article Title: Comparative Analysis of Morphological and Release Profiles in Ocular Implants of Acetazolamide Prepared by Electrospinning

doi: 10.3390/pharmaceutics13020260

Figure Lengend Snippet: FTIR spectra of blending implants compared with PCL, Lutrol F127 and acetazolamide.

Article Snippet: Acetazolamide, purity 99%, was purchased from Alfa Aesar (Heysham, UK).

Techniques:

Journal: Pharmaceutics

Article Title: Comparative Analysis of Morphological and Release Profiles in Ocular Implants of Acetazolamide Prepared by Electrospinning

doi: 10.3390/pharmaceutics13020260

Figure Lengend Snippet: Melting temperature, onset degradation temperature and variation of melting enthalpy of pure substances: acetazolamide, Lutrol F127, PCL and the membrane formulations: EDW, MD and blending.

Article Snippet: Acetazolamide, purity 99%, was purchased from Alfa Aesar (Heysham, UK).

Techniques: Membrane

Journal: Frontiers in Ecology and Evolution

Article Title: Mechanisms of carbon dioxide detection in the earthworm Dendrobaena veneta

doi: 10.3389/fevo.2023.1202410

Figure Lengend Snippet: FIGURE 7 Exudate excretion after treatment with blockers and inhibitors versus treatment with the respective vehicle. (A) The carbonic anhydrase inhibitors acetazolamide (general CA inhibitor, p<0.01, n=8-13) and indisulam (CA IX/XII inhibitor, p<0.05, n=6-8) significantly muted the exudate response by a two-way ANOVA and significant differences were detected via Tukey’s HSD test. S4 (CA IX inhibitor, p>0.05, n=5-8), U-104 (CA IX/XII inhibitor, p>0.05, n=6-8) and topiramate (CA II/IV inhibitor, p>0.05, n=6-8) did not significantly alter the response. (B) Receptor blockers amiloride (ENaC blocker, p<0.0001, n=3-8), diminazene aceturate (ASIC3 blocker, p<0.0001, n=3-8), and ruthenium red (Ca2+ channel blocker, p<0.01, n=5-13) significantly muted the exudate response by a two-way ANOVA, and significant differences were detected via Tukey’s HSD test. HC030013 (TRPA1 blocker, p>0.05, n=6-12), methylene blue (guanylate cyclase inhibitor, p>0.05, n=6-8), and ZnCl2 (OTOP1 blocker, p>0.05, n=6-8) did not significantly alter the response. Graphed values are means ± SEM; * p <0.05, ** p <0.01, *** p <0.001 via Tukey’s HSD test.

Article Snippet: Inhibitors and blockers were sourced from Tocris Bioscience (Acetazolamide, Diminazene Aceturate, S4, Topiramate) and Sigma-Aldrich (Amiloride, HC030013, Methylene Blue, Ruthenium Red, ZnCl2, U-104).

Techniques:

Journal: Frontiers in Ecology and Evolution

Article Title: Mechanisms of carbon dioxide detection in the earthworm Dendrobaena veneta

doi: 10.3389/fevo.2023.1202410

Figure Lengend Snippet: FIGURE 8 AITC induced exudate excretion after treatment with blockers. AITC significantly increased exudate production (p < 0.01, n=8) and was altered by blocker treatment (p < 0.0001, n=4) compared to control (n=4) via two-way ANOVA. The response to AITC was significantly muted by the TRPA1 blocker HC030013 (p<0.0001, n=7), 5mM amiloride (p<0.0001, n=4), 0.05mM diminazene aceturate (p<0.0001, n=5) and 0.1mM diminazene aceturate (p<001, n=5) via Tukey’s HSD test; while acetazolamide (n=4) and indisulam (n=5) did not significantly alter exudate production. Graphed values are means ± SEM; * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Inhibitors and blockers were sourced from Tocris Bioscience (Acetazolamide, Diminazene Aceturate, S4, Topiramate) and Sigma-Aldrich (Amiloride, HC030013, Methylene Blue, Ruthenium Red, ZnCl2, U-104).

Techniques: Control

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Altered hemodynamics and vascular reactivity in a mouse model with severe pericyte deficiency

doi: 10.1177/0271678X221147366

Figure Lengend Snippet: Impaired dilatory cerebral blood volume responses in Pdgfb ret/ret mice by fMRI. (a) Cortical blood volume changes to acetazolamide injection (30 mg/kg; Time 0) over time. Mean ± SD (shaded area) and (b) Area under the curve (AUC) was calculated from Time 0 until the end of scanning. Each value represents the AUC for one animal. n = 6 Control mice; n = 5 Pdgfb ret/ret mice. Dot plot is mean ± SD.

Article Snippet: After the 30th repetition (20 minutes), acetazolamide (30 mg/kg body weight; Diamox® parenteral, Goldshield Pharmaceuticals Ltd, Croydon, UK) was administered i.v. as a bolus.

Techniques: Injection, Control