Journal: Cell reports

Article Title: KLF10 Deficiency in CD4 + T Cells Triggers Obesity, Insulin Resistance, and Fatty Liver

doi: 10.1016/j.celrep.2020.108550

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: HDL colorimetric assay (HDL) , BioAssay Systems , #E2HL-100.

Techniques: Phospho-proteomics, Recombinant, Reverse Transcription, Cell Isolation, Glucose Assay, Colorimetric Assay, Bioassay, Enzyme-linked Immunosorbent Assay, Gene Expression, Transgenic Assay, Knock-Out, Software, Flow Cytometry

Journal: International Journal of Epidemiology

Article Title: Cohort Profile: The Oxford Biobank

doi: 10.1093/ije/dyx132

Figure Lengend Snippet: Baseline characteristics of the OBB participants

Article Snippet: HDL- and LDL-cholesterol , , Analysed using Randox kits adapted for use on the Ilab 600/650 analysers (Randox Laboratories, Crumlin, Northern Ireland).

Techniques: Activity Assay

Journal: International Journal of Epidemiology

Article Title: Cohort Profile: The Oxford Biobank

doi: 10.1093/ije/dyx132

Figure Lengend Snippet: List of platforms used for biochemical tests

Article Snippet: HDL- and LDL-cholesterol , , Analysed using Randox kits adapted for use on the Ilab 600/650 analysers (Randox Laboratories, Crumlin, Northern Ireland).

Techniques: Northern Blot, RIA Assay

Journal: Endocrinology

Article Title: Inhibition of Lysyl Oxidase by Cortisol Regeneration in Human Amnion: Implications for Rupture of Fetal Membranes.

doi: 10.1210/en.2016-1406

Figure Lengend Snippet: Figure 1. Changes of 11-HSD1, cortisol and LOX in human amnion tissue at parturition and reciprocal regulation of 11- HSD1 and LOX by cortisol in human amnion tissue explants. (A and B) Comparison of 11-HSD1 protein, cortisol (A) and LOX activity (B) between amnion tissues collected at term after elective Cesarean section (term nonlabor) and after vaginal delivery with spontaneous rupture of the membranes (term labor). n 7 for 11-HSD1, n 6 for LOX and n 10 for cortisol. (C) Effects of cortisol (1 M) on 11-HSD1 and LOX protein abundance in cultured human amnion tissue explants. n 3. The top panels of A and C are the representative immunoblots. Data are the mean SEM. Statistical analysis was performed using unpaired (A, B) and paired (C) Student’s t test. *P .05, **P .01 vs. nonlabor or control.

Article Snippet: Following blocking, antibodies against 11 -HSD1 (Origene, Rockville, MD), LOX (Abcam, Cambridge, UK), cytokeratin (Santa Cruz Biotechnology, Santa Cruz, CA) and vimentin (Santa Cruz) at 1:100 dilutions were applied.

Techniques: Comparison, Activity Assay, Quantitative Proteomics, Cell Culture, Western Blot, Control

Journal: Endocrinology

Article Title: Inhibition of Lysyl Oxidase by Cortisol Regeneration in Human Amnion: Implications for Rupture of Fetal Membranes.

doi: 10.1210/en.2016-1406

Figure Lengend Snippet: Figure 2. Distribution of LOX and 11-HSD1 in human fetal membranes and the effects of cortisol in human amnion epithelial and fibroblast cells. (A-C) Immunofluorescence staining of 11-HSD1 (A, orange) and LOX (B, green) in human fetal membranes. Yellow color after merge represents the colocalization of 11-HSD1 and LOX (C). Nuclei were counterstained blue with DAPI. Scale bar, 10 m. (D) Immunofluorescent staining of LOX and LOX activity in cultured amnion epithelial and fibroblast cells. The top panel is the double staining of LOX (green) and vimentin (red) in amnion fibroblasts, and LOX (green) and cytokeratin (red) in amnion epithelial cells. Nuclei were counterstained blue with DAPI. Arrows indicate amnion epithelial cells stained positive for LOX. Scale bar, 10 m. The bottom panel illustrates the effect of cortisol (1 M) on LOX activity in amnion fibroblasts and epithelial cells. n 3. E, The reciprocal effects of cortisol (1 M) on 11-HSD1 and LOX protein abundance in human amnion epithelial and fibroblast cells. (F-H) Immunofluorescent double staining of 11-HSD1 (F, orange) and LOX (G, green) in human amnion fibroblasts. Nuclei were counterstained blue with DAPI. Yellow color after merge represents the colocalization of 11- HSD1 and LOX (H). Scale bar, 10 m. (I and J) Concentration-dependent induction of 11-HSD1 (I, n 5) and reduction of LOX (J, n 5 for mRNA, n 7 for protein) by cortisol (0.01–1 M) in human amnion fibroblasts. The top panels of E, I and J are the representative immunoblots. ae: amnion epithelial cells; af: amnion fibroblasts; ct: chorion trophoblasts. Data are the mean SEM. Statistical analysis was performed with paired Student’s t test (E, K) or one-way ANOVA test (I and J). *P .05, **P .01, ***P .001 vs. control or 0 M.

Article Snippet: Following blocking, antibodies against 11 -HSD1 (Origene, Rockville, MD), LOX (Abcam, Cambridge, UK), cytokeratin (Santa Cruz Biotechnology, Santa Cruz, CA) and vimentin (Santa Cruz) at 1:100 dilutions were applied.

Techniques: Immunofluorescence, Staining, Activity Assay, Cell Culture, Double Staining, Quantitative Proteomics, Concentration Assay, Western Blot, Control

Journal: Endocrinology

Article Title: Inhibition of Lysyl Oxidase by Cortisol Regeneration in Human Amnion: Implications for Rupture of Fetal Membranes.

doi: 10.1210/en.2016-1406

Figure Lengend Snippet: Figure 3. Involvement of 11-HSD1 in the down-regulation of LOX by cortisone in human amnion fibroblasts. (A) Attenuation of cortisone (5 M)-induced reduction in LOX mRNA and protein by 11-HSD1 inhibitor CBX (1 M). n 3. (B) Attenuation of cortisone (5 M)-induced reduction in LOX mRNA and LOX protein by 11-HSD1 inhibitor 10j (1 M). n 3 for mRNA, n 4 for protein. (C) Attenuation of cortisone (5 M)-induced reduction in LOX mRNA, protein and activity by siRNA (50 nM)-mediated knock-down of 11-HSD1. n 5. The top panels of A-C are the representative immunoblots. Data are the mean SEM. Statistical analysis was performed with one-way ANOVA test. *P .05, ***P .001 vs. control or negative scrambled siRNA control; #P .05, ##P .01, ###P .001 vs. cortisone alone.

Article Snippet: Following blocking, antibodies against 11 -HSD1 (Origene, Rockville, MD), LOX (Abcam, Cambridge, UK), cytokeratin (Santa Cruz Biotechnology, Santa Cruz, CA) and vimentin (Santa Cruz) at 1:100 dilutions were applied.

Techniques: Activity Assay, Knockdown, Western Blot, Control

Journal: Endocrinology

Article Title: Inhibition of Lysyl Oxidase by Cortisol Regeneration in Human Amnion: Implications for Rupture of Fetal Membranes.

doi: 10.1210/en.2016-1406

Figure Lengend Snippet: Figure 6. Involvement of GR and nGRE in the inhibition of LOX promoter activity by cortisol and the working model illustrating LOX inhibition in human amnion fibroblasts. (A) Inhibition of LOX promoter (-850bp 126bp, -450bp 126bp) activities by cortisol (1 M). n 4. (B) Blockade of cortisol (1 M)-induced inhibition of LOX promoter (-850bp 126bp) activity by RU486 (1 M, n 4) or siRNA (50 nM, n 5)-mediated knock-down of GR. (C) Enhanced enrichment of GR in LOX promoter spanning the putative nGRE upon cortisol (1 M) stimulation of human amnion fibroblasts as measured with the ChIP assay. n 4. TSS: transcription start site, Fwd and Rev indicate the aligning position of the forward primer (Fwd) and reverse primer (Rev). (D) Abolition of cortisol (1 M)-induced inhibition of LOX promoter (-850bp 126bp) activity by introduction of nucleotide mutations (nucleotides in red) into the putative nGRE (-260 to -252bp) but not by nucleotide mutations in the adjacent CRE (-332 to -325bp) (nucleotides in red). n 3. Data are the mean SEM. Statistical analysis was performed with paired Student’s t test (A, C and D) or one-way ANOVA test (B). *P .05, **P .01 vs. control or negative scrambled siRNA control; #P .05 vs. control or cortisol alone. (E) A working model illustrating the participation of the feed-forward regeneration of cortisol by 11-HSD1 in the structural remodeling in human amnion. Toward the end of gestation, accumulation of cortisol regenerated by 11-HSD1 in the amnion not only induces 11-HSD1 expression, but also down-regulates LOX expression via GR-mediated inhibition of LOX expression in amnion fibroblasts. At the same time, accumulation of cortisol also up-regulates PGE2 production via induction of cPLA2 and COX-2 expression. Increased PGE2 production, in turn, inhibits LOX expression via EP2/EP4 receptors in amnion fibroblasts. Cortisol and PGE2 may act in a coordinated fashion to cause dramatic down-regulation of LOX expression toward the end of gestation resulting in the reduction of collagen cross-linking and weakened tensile strength of the amnion. Reduction in collagen cross-linking may also predispose it to degradation by MMP, which further reduces the tensile strength of the amnion. All of these effects may be implicated in ROM during normal parturition in humans. Preterm activation of this feed-forward mechanism may result in PPROM and preterm birth. Blue arrows indicate stimulation and red lines indicate inhibition. E, cortisone; F, cortisol.

Article Snippet: Following blocking, antibodies against 11 -HSD1 (Origene, Rockville, MD), LOX (Abcam, Cambridge, UK), cytokeratin (Santa Cruz Biotechnology, Santa Cruz, CA) and vimentin (Santa Cruz) at 1:100 dilutions were applied.

Techniques: Inhibition, Activity Assay, Knockdown, Control, Expressing, Activation Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Great Healing Potential Hidden in Plant Preparations of Antioxidant Properties: A Return to Nature?

doi: 10.1155/2020/8163868

Figure Lengend Snippet: The protective effects of plant origin substances against toxicity of ethanol.

Article Snippet: Leung et al. [ ] , Camellia sinensis O. Ktze (black tea) extract (QP-lack tea extract, Quality Phytochemicals LLC) 15 mg/kg/day, i.g., for 4 weeks, 3 months after ovariectomy Female Sprague-Dawley rats, 3 months, 200–230 g , Ovariectomy Body weight: ↑ Serum: 17 β -estradiol, cGMP ↓, non-HDL-c, HDL-c, TC, TG ↑ Aorta: peNOS ↓, NOX2, NOX4 ↑ Endothelium of aorta: ROS ↑ , Body weight: (0) Serum: TG ↓, 17 β -estradiol, non-HDL-c (0), HDL-c, TC (++), cGMP (+++) Aorta: peNOS, NOX2, NOX4 (+++) Endothelium of aorta: ROS (+++) , Not studied.

Techniques: AST Assay, Multiple Displacement Amplification, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Great Healing Potential Hidden in Plant Preparations of Antioxidant Properties: A Return to Nature?

doi: 10.1155/2020/8163868

Figure Lengend Snippet: The protective effects of plant substances against toxicity of antipyretic and analgesic drugs.

Article Snippet: Leung et al. [ ] , Camellia sinensis O. Ktze (black tea) extract (QP-lack tea extract, Quality Phytochemicals LLC) 15 mg/kg/day, i.g., for 4 weeks, 3 months after ovariectomy Female Sprague-Dawley rats, 3 months, 200–230 g , Ovariectomy Body weight: ↑ Serum: 17 β -estradiol, cGMP ↓, non-HDL-c, HDL-c, TC, TG ↑ Aorta: peNOS ↓, NOX2, NOX4 ↑ Endothelium of aorta: ROS ↑ , Body weight: (0) Serum: TG ↓, 17 β -estradiol, non-HDL-c (0), HDL-c, TC (++), cGMP (+++) Aorta: peNOS, NOX2, NOX4 (+++) Endothelium of aorta: ROS (+++) , Not studied.

Techniques: AST Assay, Multiple Displacement Amplification, Histopathology, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Great Healing Potential Hidden in Plant Preparations of Antioxidant Properties: A Return to Nature?

doi: 10.1155/2020/8163868

Figure Lengend Snippet: The protective effects of plant origin materials in the animal model of menopause.

Article Snippet: Leung et al. [ ] , Camellia sinensis O. Ktze (black tea) extract (QP-lack tea extract, Quality Phytochemicals LLC) 15 mg/kg/day, i.g., for 4 weeks, 3 months after ovariectomy Female Sprague-Dawley rats, 3 months, 200–230 g , Ovariectomy Body weight: ↑ Serum: 17 β -estradiol, cGMP ↓, non-HDL-c, HDL-c, TC, TG ↑ Aorta: peNOS ↓, NOX2, NOX4 ↑ Endothelium of aorta: ROS ↑ , Body weight: (0) Serum: TG ↓, 17 β -estradiol, non-HDL-c (0), HDL-c, TC (++), cGMP (+++) Aorta: peNOS, NOX2, NOX4 (+++) Endothelium of aorta: ROS (+++) , Not studied.

Techniques: Animal Model

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Great Healing Potential Hidden in Plant Preparations of Antioxidant Properties: A Return to Nature?

doi: 10.1155/2020/8163868

Figure Lengend Snippet: The protective effects of plant origin materials in animal models of obesity.

Article Snippet: Leung et al. [ ] , Camellia sinensis O. Ktze (black tea) extract (QP-lack tea extract, Quality Phytochemicals LLC) 15 mg/kg/day, i.g., for 4 weeks, 3 months after ovariectomy Female Sprague-Dawley rats, 3 months, 200–230 g , Ovariectomy Body weight: ↑ Serum: 17 β -estradiol, cGMP ↓, non-HDL-c, HDL-c, TC, TG ↑ Aorta: peNOS ↓, NOX2, NOX4 ↑ Endothelium of aorta: ROS ↑ , Body weight: (0) Serum: TG ↓, 17 β -estradiol, non-HDL-c (0), HDL-c, TC (++), cGMP (+++) Aorta: peNOS, NOX2, NOX4 (+++) Endothelium of aorta: ROS (+++) , Not studied.

Techniques: Expressing, Multiple Displacement Amplification, AST Assay

Journal: Scientific Reports

Article Title: Active site competition is the mechanism for the inhibition of lipoprotein-associated phospholipase A 2 by detergent micelles or lipoproteins and for the efficacy reduction of darapladib

doi: 10.1038/s41598-020-74236-0

Figure Lengend Snippet: Effects of detergents, latex nanobeads and lipoprotein particles on the activation and inhibition of Lp-PLA 2 . ( a ) Lp-PLA 2 activity in TBS, pH 7.4, containing 10 µM 14:0 NPS-PC were followed with the titration of CHAPS, Triton X-100, digitonin and Tween-20 respectively as described in Experimental Procedures. ( b ) Lp-PLA 2 activity in 540 µM 14:0 NPS-PC were followed with the titration of CHAPS, digitonin, Triton X-100 and SNS respectively as described in Experimental Procedures. ( c ) Lineweaver–Burk plots for competitive inhibition of Lp-PLA 2 by different concentrations of Tween-20 (TW20) as indicated. ( d ) Inhibition of Lp-PLA 2 by darapladib in presence of 80 µM and 800 µM of Tween-20. Assay was carried out in 100 mM HEPES buffer, pH 7.5, containing 0.54 mM 14:0 NPS-PC, 4 mM EDTA and 10 mM SNS. ( e ) Lp-PLA 2 activity in TBS, pH 7.4, containing 10 µM 14:0 NPS-PC were followed with the titration of four different polystyrene nanobeads. ( f ) Lp-PLA 2 activity in TBS, pH 7.4, containing 10 µM 14:0 NPS-PC were followed with the titration of Lp-PLA 2 -depleted HDL and LDL as indicated.

Article Snippet: Concentrated human LDL and HDL were purchased from Lee BioSolutions in St. Louis.

Techniques: Activation Assay, Inhibition, Activity Assay, Titration

Journal: Scientific Reports

Article Title: Active site competition is the mechanism for the inhibition of lipoprotein-associated phospholipase A 2 by detergent micelles or lipoproteins and for the efficacy reduction of darapladib

doi: 10.1038/s41598-020-74236-0

Figure Lengend Snippet: Effects of detergents on the activity of Lp-PLA 2 associated with LDL and HDL. ( a ) Changes of LDL Lp-PLA 2 activity with titration of detergents. ( b ) Changes of HDL Lp-PLA 2 activity with titration of detergents. Lp-PLA 2 activity of 20 µl LDL at 1 mg/ml or HDL at 0.15 mg/ml cholesterol were determined in final volume of 130 µl containing 10 µM of 14:0 NPS-PC and detergents as indicated.

Article Snippet: Concentrated human LDL and HDL were purchased from Lee BioSolutions in St. Louis.

Techniques: Activity Assay, Titration

Journal: Scientific Reports

Article Title: Active site competition is the mechanism for the inhibition of lipoprotein-associated phospholipase A 2 by detergent micelles or lipoproteins and for the efficacy reduction of darapladib

doi: 10.1038/s41598-020-74236-0

Figure Lengend Snippet: The relationship between Lp-PLA 2 activity and lipoprotein concentration. ( a ) Lp-PLA 2 activity by incrementation of lipoprotein. Lp-PLA 2 activity of 20 µl lipoproteins (LDL and HDL) were determined in 10 µM of 14:0 NPS-PC with increasing concentration of lipoproteins as indicated. Lipoprotein concentration is expressed as cholesterol content. ( b ) Inhibition of Lp-PLA 2 activity by Lp-PLA 2 -depleted lipoproteins. Lp-PLA 2 activity of 20 µl LDL at 0.4 mg/ml or HDL at 0.04 mg/ml were determined in 10 µM of 14:0 NPS-PC with increasing concentrations of Lp-PLA 2 -depleted lipoproteins as indicated. Lipoprotein concentration is expressed as cholesterol content. Lp-PLA 2 in lipoproteins was depleted by incubation with Pefabloc SC as descripted in Experimental Procedure.

Article Snippet: Concentrated human LDL and HDL were purchased from Lee BioSolutions in St. Louis.

Techniques: Activity Assay, Concentration Assay, Inhibition, Incubation

Journal: Scientific Reports

Article Title: Active site competition is the mechanism for the inhibition of lipoprotein-associated phospholipase A 2 by detergent micelles or lipoproteins and for the efficacy reduction of darapladib

doi: 10.1038/s41598-020-74236-0

Figure Lengend Snippet: Inhibition of Lp-PLA 2 activity associated with LDL and HDL by darapladib. ( a ) Inhibition of LDL Lp-PLA 2 activity under three different conditions: (1) no preincubation with darapladib, (2) preincubated with darapladib for 30 min at ambient temperature and (3) preincubated with darapladib for 30 min at ambient temperature in the presence of 40 mM CHAPS. ( b ) Inhibition of HDL Lp-PLA 2 activity under the same conditions as in (a). ( c ) Comparing the inhibition of Lp-PLA 2 activity associated with LDL and HDL by darapladib under different concentration of lipoproteins as indicated without preincubation. ( d ) Comparing the inhibition of Lp-PLA 2 activity associated with LDL and HDL by darapladib under different concentration of lipoproteins as indicated with preincubation at ambient temperature for 30 min before adding 14:0 NPS-PC. ( e ) Comparing the inhibition of Lp-PLA 2 activity associated with LDL and HDL by darapladib under different concentration of lipoproteins as indicated with preincubation in the presence of 40 mM CHAPS. ( f ) Fractionation of purified HDL and LDL by TSKgel G3000SW XL column in TBS, pH 7.4. Forty µl of 1.0 mg/ml lipoproteins were injected. Fractions were assayed for Lp-PLA 2 activity as described in Experimental Procedures. ( g ) Fractionation of human serum with and without treatment with 150 µM darapladib under the conditions as indicated by TSKgel G3000SW XL column in TBS, pH 7.4. Forty µl of neat serum were injected. Fractions were assayed for residual Lp-PLA 2 activity as described in Experimental Procedures.

Article Snippet: Concentrated human LDL and HDL were purchased from Lee BioSolutions in St. Louis.

Techniques: Inhibition, Activity Assay, Concentration Assay, Fractionation, Purification, Injection

Journal: Scientific Reports

Article Title: Active site competition is the mechanism for the inhibition of lipoprotein-associated phospholipase A 2 by detergent micelles or lipoproteins and for the efficacy reduction of darapladib

doi: 10.1038/s41598-020-74236-0

Figure Lengend Snippet: Transport of spiked recombinant Lp-PLA 2 (rLp-PLA 2 ) between purified HDL and LDL. Endogenous Lp-PLA 2 were depleted from purified LDL and HDL by incubation with magnetic beads coupled with anti-Lp-PLA 2 antibodies and separation of the supernatants. The Lp-PLA 2 depleted LDL and HDL (LDL-0 and HDL-0) were spiked in duplication with 25 µg/ml of rLp-PLA 2 and incubated at ambient temperature for 16 h (LDL-1, LDL-2, HDL-1 and HDL-2). LDL and HDL spiked with Lp-PLA 2 were further mixed with blank HDL (HDL-0) or LDL (LDL-0) respectively as shown. All mixtures were further incubated at ambient temperature for additional 6 h and stored in 2–8 ºC for 16 h before subjected to resolution by non-denatured gel electrophoresis as described in Experimental Procedures.

Article Snippet: Concentrated human LDL and HDL were purchased from Lee BioSolutions in St. Louis.

Techniques: Recombinant, Purification, Incubation, Magnetic Beads, Nucleic Acid Electrophoresis

Journal: Scientific Reports

Article Title: Active site competition is the mechanism for the inhibition of lipoprotein-associated phospholipase A 2 by detergent micelles or lipoproteins and for the efficacy reduction of darapladib

doi: 10.1038/s41598-020-74236-0

Figure Lengend Snippet: Distribution of 14:0 NPS-PC and darapladib under different conditions. Assay methods are described in Experimental Procedures. ( a ) Distribution of 14:0 NPS-PC in TBS, pH 7.4, with and without detergents. Twenty µl of 14:0 NPS-PC at 10 µM were resolved by a TSKgel G3000SW XL column in TBS, pH 7.4, containing no detergent, 10 mM CHAPS or 60 µM Tween-20. Absorbance at 260 nm were monitored for 14:0 NPS-PC. Bulk phase is after fraction number 17 or after elution time 17 min. ( b ) Distribution of darapladib in TBS, pH 7.4, with and without detergents. Forty µl of darapladib in mobile phase buffer at 0.2 µM were resolved by a TSKgel G3000SW XL column in TBS, pH 7.4, containing no detergent, 10 mM CHAPS or 600 µM Tween-20. Relative darapladib concentration in each fraction was shown by inhibition of spiked Lp-PLA 2 . ( c ) Distribution of darapladib in TBS, pH 7.4, containing HDL or LDL. Forty µl of 1 mg/ml HDL or LDL with or without 0.2 µM darapladib were resolved by a TSKgel G3000SW XL column in TBS, pH 7.4. Relative darapladib concentration in each fraction was quantitated by inhibition of spiked Lp-PLA 2 . ( d ) Distribution of darapladib in human serum. Forty µl of neat human serum with or without 150 µM darapladib under the conditions as indicated were resolved by a TSKgel G3000SW XL column in TBS, pH 7.4. Relative darapladib concentration in each fraction was determined by inhibition of spiked Lp-PLA 2 .

Article Snippet: Concentrated human LDL and HDL were purchased from Lee BioSolutions in St. Louis.

Techniques: Concentration Assay, Inhibition

Journal: Journal of Clinical Medicine

Article Title: Associations between High-Density Lipoprotein Functionality and Major Adverse Cardiovascular Events in Patients Who Have Undergone Coronary Computed Tomography Angiography

doi: 10.3390/jcm10112431

Figure Lengend Snippet: ( A ) Relative caspase 3/7 activity, ( B ) relative total caspase 3/7 activity, ( C ) relative secretion of MCP-1 and ( D ) relative total secretion of MCP-1 in the MACE(+) and MACE(−) groups. A.U : arbitrary unit.

Article Snippet: We analyzed the HDL-suppressed caspase 3/7 activity with an ex vivo system that used the H9C2 cell line of embryonic rat cardiomyoblasts (ATCC ® , CRL-1446 T , Manassas, VA, USA) and apo-B-depleted plasma from the study participants as samples.

Techniques: Activity Assay